RESUMEN

The exact aetiology of the cause of death (COD) remains unknown for a high proportion of deaths caused by infectious diseases. Complete diagnostic autopsy (CDA) is considered the gold standard to determine COD, but it is often not used in low and middle-income countries (LMIC), including Vietnam, for a variety of reasons. One alternative is minimally invasive tissue sampling (MITS). This study was part of a larger project to explore the perceptions of MITS in Vietnam to provide recommendations for its potential implementation. We collected in-depth interviews and focus group discussions that explored MITS acceptability from 96 participants, including key informants, healthcare workers, community stakeholders, and people who had family members pass away recently. Participants highlighted the minimally invasive nature as the most considerable strength of MITS that could make it acceptable in Vietnam compared with CDA. However, participants still had concerns including the accuracy of the technique, the potential damage on the body, and the potential conflicts MITS results could have with original clinical diagnoses. Overall, the hypothetical acceptability of MITS was not clear-cut. It is imperative to consider acceptability before implementing new medical procedures into a given context, especially when it revolves around death, dying and the body.

Asunto(s)

Autopsia , Grupos Focales , Entrevistas como Asunto , Investigación Cualitativa , Humanos , Vietnam , Femenino , Masculino , Adulto , Autopsia/métodos , Causas de Muerte , Persona de Mediana Edad , Manejo de Especímenes

RESUMEN

With the continuous advancements in precision medicine and the relentless pursuit of minimally invasive techniques, Natural Orifice Specimen Extraction Surgery (NOSES) has emerged. Compared to traditional surgical methods, NOSES better embodies the principles of minimally invasive surgery, making scar-free operations possible. In recent years, with the progress of science and technology, Robot-Assisted Laparoscopic Surgery has been widely applied in the treatment of colorectal cancer. Robotic surgical systems, with their clear surgical view and high operational precision, have shown significant advantages in the treatment process. To further improve the therapeutic outcomes for colorectal cancer patients, some scholars have attempted to combine robotic technology with NOSES. However, like traditional open surgery or laparoscopic surgery, the use of the robotic platform presents both advantages and limitations. Therefore, this study reviews the current research status, progress, and controversies regarding Robot-Assisted Laparoscopic Natural Orifice Specimen Extraction Surgery for colorectal cancer, aiming to provide clinicians with more options in the diagnosis and treatment of colorectal cancer.

Asunto(s)

Neoplasias Colorrectales , Laparoscopía , Cirugía Endoscópica por Orificios Naturales , Procedimientos Quirúrgicos Robotizados , Humanos , Neoplasias Colorrectales/cirugía , Procedimientos Quirúrgicos Robotizados/métodos , Cirugía Endoscópica por Orificios Naturales/métodos , Laparoscopía/métodos , Predicción , Manejo de Especímenes/métodos

RESUMEN

The molecular reverse transcription-polymerase chain reaction (RT-PCR) testing of respiratory tract swabs has become mandatory to confirm the diagnosis of coronavirus disease 2019 (COVID-19). However, RT-PCR tests are expensive, require standardized equipment, and relatively long testing times, and the sample pooling method has been introduced to solve this issue. The aim of this study was to compare the cycle threshold (Ct) values of the individual sample and pooled sample methods to assess how accurate the pooling method was. Repeat RT-PCR examinations were initially performed to confirm the Ct values for each sample before running the pooled test procedure. Sample extraction and amplification were performed in both assays to detect ORF1ab, N, and E genes with a cut-off point value of Ct <38. Overall, there was no difference in Ct values between individual sample and pooled sample groups at all concentrations (p=0.259) and for all pooled sizes. Only pooled size of five could detect the Ct value in the pooled samples for all concentration samples, including low-concentration sample (Ct values 36 to 38). This study highlighted that pooled RT-PCR testing strategy did not reduce the quality of individually measured RT-PCR Ct values. A pool size of five could provide a practical technique to expand the screening capacity of RT-PCR.

Asunto(s)

COVID-19 , Nasofaringe , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , SARS-CoV-2 , Manejo de Especímenes , Humanos , Nasofaringe/virología , SARS-CoV-2/genética , COVID-19/diagnóstico , COVID-19/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Manejo de Especímenes/métodos , Prueba de Ácido Nucleico para COVID-19/métodos

RESUMEN

INTRODUCTION: The human gut microbiota has the potential to modulate the outcomes of several human diseases. This effect is likely to be mediated through interaction with the host immune system. This protocol details the establishment of a biorepository of clinically annotated samples, which we will use to explore correlations between the gut microbiota and the immune system of immune-compromised patients. We aim to identify microbiome-related risk factors for adverse outcomes. METHODS AND ANALYSES: This is a protocol for the development of a biorepository of clinically annotated samples collected prospectively across three centres in Melbourne, Australia. Participants will be recruited across the following clinical streams: (1) acute leukaemia and allogeneic stem cell transplant; (2) end-stage liver disease and liver transplant; (3) patients receiving any cancer immunotherapies (eg, chimeric antigen receptor therapy); (4) deceased organ donors and (5) healthy adult controls. Participants will be asked to provide paired peripheral blood and microbiota samples (stool and saliva) at either (1) single time point for healthy controls and deceased organ donors or (2) longitudinally over multiple prespecified or event-driven time points for the remaining cohorts. Sampling of fluid from bronchoalveolar lavage and colonoscopy or biopsy of tissues undertaken during routine care will also be performed. ETHICS AND DISSEMINATION: Ethical approval has been obtained from the relevant local ethics committee (The Royal Melbourne Hospital Human Research Ethics Committee). The results of this study will be disseminated by various scientific platforms including social media, international presentations and publication in peer-reviewed journals. TRIAL REGISTRATION NUMBER: ACTRN12623001105639. Date registered 20 October 2023.

Asunto(s)

Microbioma Gastrointestinal , Humanos , Australia , Interacciones Microbiota-Huesped/inmunología , Bancos de Muestras Biológicas , Estudios Prospectivos , Proyectos de Investigación , Manejo de Especímenes/métodos

RESUMEN

Intimate partner violence affects 20-30% of women in the United States. Disparities in routine cervical cancer surveillance have been demonstrated in certain populations, including victims of intimate partner violence (IPV). This study examined and assessed the acceptability of high-risk HPV (hrHPV) self-collection among individuals who have experienced IPV. We conducted an observational study using qualitative data collection and analysis. We interviewed individuals with a history of IPV and who currently reside in Oregon. This study identified key themes describing knowledge and attitudes towards cervical cancer screening for individuals who have experienced IPV. They include: guideline knowledge, prior office-based cervical cancer screening experience, barriers to cervical cancer screening, at-home hrHPV self-collection experience, and testing confidence. Participants experienced fewer barriers and expressed increased comfort and control with hrHPV self-collection process. Individuals with a history of IPV have lower rates of cervical cancer screening adherence and higher rates of cervical dysplasia and cancer than other populations. The patient-centered approach of hrHPV self-collection for cervical cancer screening can reduce barriers related to the pelvic exam and empower patients to reduce their risks of developing cervical cancer by enabling greater control of the testing process.

Asunto(s)

Detección Precoz del Cáncer , Investigación Cualitativa , Neoplasias del Cuello Uterino , Humanos , Femenino , Neoplasias del Cuello Uterino/diagnóstico , Adulto , Detección Precoz del Cáncer/métodos , Detección Precoz del Cáncer/psicología , Persona de Mediana Edad , Infecciones por Papillomavirus/diagnóstico , Conocimientos, Actitudes y Práctica en Salud , Violencia de Pareja/psicología , Violencia de Pareja/estadística & datos numéricos , Manejo de Especímenes/métodos , Oregon , Autocuidado/métodos , Autocuidado/psicología , Sobrevivientes/psicología , Frotis Vaginal/métodos , Frotis Vaginal/psicología , Aceptación de la Atención de Salud/psicología , Aceptación de la Atención de Salud/estadística & datos numéricos , Adulto Joven

RESUMEN

BACKGROUND: The detection of causative pathogens plays a crucial role in the diagnosis and targeted treatment of periprosthetic joint infections (PJI). While there have been improvements in analytic methods in the past, pre-analytical procedures have not yet been sufficiently investigated. The objective of this study was to compare the culture yield of four different pre-analytical procedures. METHODS: Patients with perioperative diagnosis of PJI were included in a single center cross-sectional study (2021-2022). Tissue samples (n = 20) of each patient were randomly and equally distributed to each of the four study arms. Tissue samples were either send to the laboratory without culture medium (group A) or were transported in thioglycolate medium immediately after sampling at three different temperatures (room temperature, 4 °C, 37° for 24 h; group B-D). Culture media were investigated for growth on days 1, 3, 7, 12, 14. All organisms, the number of positive samples and the time to positivity were recorded and compared between the study arms. Single positive cultures were considered as contamination. RESULTS: In total, 71 patients were included. The proportions of culture negative samples (10-15%) and polymicrobial infections (51-54%) were comparable between the four arms. Seven patients (10%) were culture-negative in group A, but showed growth in thioglycolate media (group B-D). Furthermore, 13% of patients showed growth in all groups, but additional organisms were cultured in thioglycolate. There was growth beyond day 7 of culturing only in thioglycolate, but not in group A. A storage temperature of 4 °C showed a longer time to positivity compared to the other groups (p < 0.001). CONCLUSIONS: Pre-analytical storage of tissue samples in thioglycolate broth did not improve the culture yield and did not detect additional cases of infection compared to the standard (pre-analytical storage in sterile containers). However, including a thioglycolate medium to the sampling algorithm reduced the rate of culture-negative infections and helped to identify additional organisms.

Asunto(s)

Medios de Cultivo , Infecciones Relacionadas con Prótesis , Humanos , Infecciones Relacionadas con Prótesis/microbiología , Infecciones Relacionadas con Prótesis/diagnóstico , Femenino , Masculino , Anciano , Estudios Transversales , Persona de Mediana Edad , Medios de Cultivo/química , Manejo de Especímenes/métodos , Bacterias/aislamiento & purificación , Bacterias/crecimiento & desarrollo , Bacterias/clasificación , Anciano de 80 o más Años , Técnicas Microbiológicas/métodos , Técnicas Bacteriológicas/métodos

RESUMEN

BACKGROUND: Human papillomavirus (HPV) self-sampling is recognized as a feasible option for enhancing screening for cervical cancer, particularly among hard-to-reach women. The magnitude of the effectiveness of screening participation under different invitation strategies was reported. This review seeks to compare the effectiveness of invitation strategies in increasing screening participation of HPV self-sampling across diverse study settings. METHODS: A systematic literature search was conducted in Embase, MEDLINE, and PubMed in April 2023. Articles were included if (1) their target participants were aged between 25 and 70 years; (2) participants in the intervention arm were randomized to receive HPV self-sampling devices through various invitation strategies; (3) participants in the control arm who either received invitations for cervical cancer screening other than HPV self-sampling or opportunistic screening as usual care; (4) studies that provided sufficient data on screening participation in HPV self-sampling as outcome measured. The study design of the included articles was limited to randomized controlled trials. RESULTS: A total of 15 articles were included in this review. Invitation strategies of disseminating HPV self-sampling devices included opt-out and opt-in. Meta-analysis revealed screening participation in the self-sampling group was significantly greater than control arm (OR 3.43, 95% CI 1.59-7.38), irrespective of the invitation strategy employed. Among invitation strategies, opt-out appeared to be more effective on increasing screening participation, compared to control and opt-in strategy (opt-out vs. control OR 3.91, 95% CI 1.82-8.42; opt-in vs. control OR 1.34, 95% CI 0.28-6.39). CONCLUSIONS: Opt-out strategy is more successful at improving screening participation compared to opt-in and routine invitation to cervical screening. It is therefore a promising way to improve participation in cervical cancer screening. The findings of this review provide important inputs to optimize strategies for inviting women to participate in vaginal HPV self-sampling across the study setting, thus improving participation in cervical cancer screening.

Asunto(s)

Detección Precoz del Cáncer , Infecciones por Papillomavirus , Ensayos Clínicos Controlados Aleatorios como Asunto , Neoplasias del Cuello Uterino , Humanos , Femenino , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/prevención & control , Detección Precoz del Cáncer/métodos , Infecciones por Papillomavirus/diagnóstico , Adulto , Persona de Mediana Edad , Manejo de Especímenes/métodos , Anciano , Aceptación de la Atención de Salud/estadística & datos numéricos , Papillomaviridae/aislamiento & purificación , Autocuidado/métodos

RESUMEN

Introduction: Human papillomavirus (HPV) testing as a method of cervical cancer screening can be performed by healthcare providers or by patients through self-sampling directly in the community, removing several barriers experienced by under screened populations. The objective of this scoping review was to determine which HPV self-sampling implementation and engagement strategies have been used to engage under screened populations (i.e., Indigenous, newcomer, and rural and remote communities) in cervical cancer screening. Methods: A scoping review was conducted searching MEDLINE, CINAHL, EMBASE, Cochrane Library, and SocINDEX from inception to August 2023. The inclusion criteria were: (1) Indigenous, newcomer, and rural and remote communities; (2) countries identified as members of the Organization for Economic Co-operation and Development; and (3) intervention included HPV self-sampling. The review was registered prior to conducting the search (https://osf.io/zfvp9). Results: A total of 26 studies out of 2,741 studies met the inclusion criteria. In-person engagement with trusted community leaders was the most widely used and accepted recruitment and engagement strategy across all three populations. Six out of seven studies with Indigenous communities distributed HPV self-sampling kits to eligible participants in person in a clinical setting for collection on site or at home. Similarly, nine of the identified studies that engaged newcomers recruited participants in person through the community, where eligible participants were either given a kit (n = 7) or received one in the mail (n = 2). Lastly, of the 10 identified studies engaging rural and remote participants in HPV self-sampling, six recruited eligible participants in person at various community locations and four used electronic medical records or registries to identify and mail kits to participants. Discussion: HPV self-sampling through in person kit distribution and mail out of HPV self-sampling kits is an effective way to increase participation rates amongst under screened populations.

Asunto(s)

Detección Precoz del Cáncer , Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Humanos , Neoplasias del Cuello Uterino/diagnóstico , Femenino , Detección Precoz del Cáncer/métodos , Infecciones por Papillomavirus/diagnóstico , Manejo de Especímenes/métodos , Tamizaje Masivo/métodos , Papillomaviridae/aislamiento & purificación , Adulto

RESUMEN

Within the field of clinical research, reports on the stability of avian serum amyloid A (SAA) under varying storage conditions are currently scarce. In this study, avian plasma samples were evaluated for SAA, a major acute-phase protein in birds, to assess how varying storage periods and repeated freeze-thaw cycles impact the stability of SAA in the frozen samples. Seven plasma samples from two species and six plasma samples from three species stored at ‒20 °C were used to evaluate the time and temperature effects accordingly. A chicken-specific SAA ELISA kit was used for the measurements. Statistical analysis was performed using SPSS, and the Kruskal-Wallis test and Spearman's correlation coefficient were applied, with statistical significance set at P < 0.05. The SAA concentrations measured daily for 30 days showed no statistically significant differences over time. Freezing-thawing was repeated five times, and a significant negative relationship was confirmed over the cycles (r=‒0.8857, P < 0.05). Although no significance was observed between a decreased concentration and the number of cycles, a decrease in the concentration of > 10% was observed after the fourth cycle in four out of six samples. However, one to three freeze-thaw cycles did not result in a significant decline. Taken together, the results indicate that a negative correlation existed between the mean concentration and multiple freeze-thaw cycles, indicating that these should be avoided where possible.

Asunto(s)

Pollos , Congelación , Proteína Amiloide A Sérica , Animales , Proteína Amiloide A Sérica/análisis , Proteína Amiloide A Sérica/metabolismo , Pollos/sangre , Factores de Tiempo , Manejo de Especímenes/veterinaria , Estabilidad Proteica , Aves/sangre

RESUMEN

BACK GROUND: The sample collection and preservation before transportation to a Forensic Science Laboratory in a medicolegal case in India is a crucial step in establishing a link between accused, victim, crime scene and the weapon. This paper discusses pioneering advancement in health technology-a state-of-the-art sample dryer developed to preserve biological samples during Forensic investigations. This unique design has been officially registered with the design office in India as a copy right. AIMS AND OBJECTIVES: The primary goal of the sample dryer is to augment the preservation of biological evidence by removing moisture content through air drying before packaging, sealing and its transportation to a Forensic Science Laboratory. MATERIAL AND METHODS: The process of air drying of biological samples by variety of experiments conducted in sample's dryer designed for the purpose which employs a range of air movement techniques including horizontal, vertical, and circular laminar flows to swiftly extract moisture from the samples. The drying duration for each experimental sample was documented upon manual confirmation of complete drying. RESULTS: The results were extraordinary, showcasing that the sample's dryer reduced the drying time drastically by up to 27 times lesser in comparison to conventional methods for identical samples. The diverse airflow patterns generated by the sample dryer unequivocally demonstrated it's utility in achieving superior sample preservation through efficient air drying. CONCLUSION: Given the exceptional outcomes observed in the experimental phase, the authors strongly advocate for the widespread adoption of this innovative sample's dryer equipment to ensure effective air drying of biological samples and their efficient sample preservation to enable successful forensic analysis as expected.

Asunto(s)

Desecación , Manejo de Especímenes , Humanos , Manejo de Especímenes/métodos , Manejo de Especímenes/instrumentación , Diseño de Equipo , Animales , Ciencias Forenses/métodos

RESUMEN

BACKGROUND: Techniques for diagnosing intestinal parasites need technological advancements in the preanalytical (collection/processing) and analytical (detection) stages. The dissolved air flotation (DAF) technique effectively recovers parasites from processed feces for routine diagnosis. Artificial intelligence (AI) is a practical and affordable alternative to modernize the analysis stage of microscopy images and generates high efficiency in the parasitological examination of feces. METHODS: The objective of this study was to standardize a laboratory protocol for stool processing using the DAF technique in conjunction with an automated diagnosis of intestinal parasites (DAPI) system. A total of 400 samples were obtained to perform the tests with the use of DAF to verify the recovery of the parasites as a function of the chemical reagent (polymer and surfactant), the volume of the flotation tube, and standardization of smear assembly on a microscopy slide, with automated analysis by DAPI. The DAF protocol that obtained the most satisfactory results in terms of parasite recovery (P < 0.05) and slide positivity was compared with the Three Fecal Test (TF-Test) protocol with manual (microscopists) and automated (DAPI) evaluation. We compared the sensitivity with the modified TF-Test technical protocol and the diagnostic agreement with the gold standard (Kappa) result. RESULTS: There was no significant difference in the parasite recovery between the 10 ml and 50 ml tubes (P > 0.05). The surfactants showed a range of parasite recoveries between 41.9% and 91.2% in the float supernatant. We obtained a maximum positivity of 73% of the assembled slides when we applied DAF processing with 7% CTAB surfactant and 57% positivity with the modified TF-Test technique. Regarding diagnostic performance, the TF-Test-modified and DAF techniques used in fecal processing for subsequent computerized analysis by AI presented sensitivities of 86% and 94%, with kappa agreements of 0.62 and 0.80 (substantial), respectively. CONCLUSIONS: The DAF protocol defined in this study and the DAPI system are innovative processes for parasite recovery and fecal debris elimination that are favorable for effectively detecting pathogenic structures in laboratory diagnosis.

Asunto(s)

Heces , Parasitosis Intestinales , Heces/parasitología , Humanos , Parasitosis Intestinales/diagnóstico , Parasitosis Intestinales/parasitología , Manejo de Especímenes/métodos , Animales , Sensibilidad y Especificidad , Automatización de Laboratorios/métodos

RESUMEN

RT-PCR is the gold standard for diagnosis of COVID-19. All RT-PCR kits are based on RNA extraction from the clinical sample. There was a sudden increase in demand of these kits, both RNA extraction and COVID-19 RT-PCR kits during the pandemic. This sudden spurt in global demand created a situation of shortage of consumables, especially the RNA extraction kits. Hence, this study was carried out to evaluate and compare COVID-19 RT-PCR without RNA extraction step using buffer R3. Sensitivity, specificity and accuracy of RT-PCR kit without RNA extraction were 89.16 %, 100% and 89.6% respectively. This approach saved more than 50 % time compared to the RT-PCR kit with RNA extraction approach allowing enhanced daily sample processing capability. RT-PCR kit without RNA extraction help in managing a greater number of samples, reduces cost and turnaround time.

Asunto(s)

COVID-19 , ARN Viral , Reacción en Cadena en Tiempo Real de la Polimerasa , SARS-CoV-2 , Sensibilidad y Especificidad , Humanos , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , COVID-19/diagnóstico , COVID-19/virología , ARN Viral/genética , ARN Viral/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Juego de Reactivos para Diagnóstico/normas , Prueba de Ácido Nucleico para COVID-19/métodos , Manejo de Especímenes/métodos

RESUMEN

BACKGROUND: Tuberculosis (TB), a major cause of disease and antimicrobial resistance, is spread via aerosols. Aerosols have diagnostic potential and airborne-microbes other than Mycobacterium tuberculosis complex (MTBC) may influence transmission. We evaluated whether PneumoniaCheck (PMC), a commercial aerosol collection device, captures MTBC and the aeromicrobiome of people with TB. METHODS: PMC was done in sputum culture-positive people (≥ 30 forced coughs each, n = 16) pre-treatment and PMC air reservoir (bag, corresponding to upper airways) and filter (lower airways) washes underwent Xpert MTB/RIF Ultra (Ultra) and 16S rRNA gene sequencing (sequencing also done on sputum). In a subset (n = 6), PMC microbiota (bag, filter) was compared to oral washes and bronchoalveolar lavage fluid (BALF). FINDINGS: 54% (7/13) bags and 46% (6/14) filters were Ultra-positive. Sequencing read counts and microbial diversity did not differ across bags, filters, and sputum. However, microbial composition in bags (Sphingobium-, Corynebacterium-, Novosphingobium-enriched) and filters (Mycobacterium-, Sphingobium-, Corynebacterium-enriched) each differed vs. sputum. Furthermore, sequencing only detected Mycobacterium in bags and filters but not sputum. In the subset, bag and filter microbial diversity did not differ vs. oral washes or BALF but microbial composition differed. Bags vs. BALF were Sphingobium-enriched and Mycobacterium-, Streptococcus-, and Anaerosinus-depleted (Anaerosinus also depleted in filters vs. BALF). Compared to BALF, none of the aerosol-enriched taxa were enriched in oral washes or sputum. INTERPRETATION: PMC captures aerosols with Ultra-detectable MTBC and MTBC is more detectable in aerosols than sputum by sequencing. The aeromicrobiome is distinct from sputum, oral washes and BALF and contains differentially-enriched lower respiratory tract microbes.

Asunto(s)

Aerosoles , Líquido del Lavado Bronquioalveolar , Tos , Mycobacterium tuberculosis , ARN Ribosómico 16S , Esputo , Humanos , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/aislamiento & purificación , Aerosoles/análisis , Esputo/microbiología , Tos/microbiología , Masculino , ARN Ribosómico 16S/genética , Adulto , Femenino , Líquido del Lavado Bronquioalveolar/microbiología , Persona de Mediana Edad , Microbiota , Microbiología del Aire , Tuberculosis Pulmonar/diagnóstico , Tuberculosis Pulmonar/microbiología , Tuberculosis/diagnóstico , Tuberculosis/microbiología , Manejo de Especímenes/métodos , Manejo de Especímenes/instrumentación , Anciano , Adulto Joven

RESUMEN

This study comprehensively evaluated the DNA/RNA Defend Pro (DRDP) sample collection buffer, designed to inactivate and stabilize patient samples. The primary objectives were to assess DRDP's efficacy in ensuring sample stability, facilitating extraction-free polymerase chain reaction (PCR), and ensuring compatibility with rapid antigen testing (RAT). Ninety-five diagnostic nasopharyngeal swab samples tested for influenza virus (influenza A), respiratory syncytial virus (RSV A), and/or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were 10-fold diluted with DRDP and anonymized. Initial characterization and retesting of these samples using cobas Liat confirmed 88 samples as positive, validating the presence of viral targets. Results from rapid antigen testing showed lower sensitivity compared to nucleic acid amplification testing (NAAT) but maintained perfect specificity, with 40 out of 88 positive samples by cobas Liat also testing positive for RAT. Direct RT-qPCR of DRDP-diluted samples demonstrated robust compatibility, with 72 out of 88 samples positive for cobas Liat also testing positive by direct RT-qPCR. Non-concordant results could be explained by the 200-fold lower input of extraction-free NAAT. Stability testing involved incubating 31 positive samples at 4 °C, 20 °C, and 37 °C for 7 days, with extraction-free NAAT. DRDP guaranteed viral RNA stability at all temperatures for influenza A, SARS-CoV-2, and RSV A, showing stability up to 7 days at 4 °C. In conclusion, DRDP is an effective stabilizing medium compatible with direct RT-qPCR and rapid antigen testing and shows great potential for optimizing diagnostic processes, particularly in resource-limited or time-sensitive scenarios.

Asunto(s)

Nasofaringe , SARS-CoV-2 , Manejo de Especímenes , Nasofaringe/virología , Humanos , Manejo de Especímenes/métodos , SARS-CoV-2/aislamiento & purificación , SARS-CoV-2/inmunología , SARS-CoV-2/genética , ARN Viral/análisis , ARN Viral/aislamiento & purificación , ARN Viral/genética , Tampones (Química) , Virus de la Influenza A/aislamiento & purificación , Virus de la Influenza A/inmunología , Virus de la Influenza A/genética , Sensibilidad y Especificidad , COVID-19/diagnóstico , COVID-19/virología , Antígenos Virales/análisis , Gripe Humana/diagnóstico , Gripe Humana/virología

RESUMEN

Of all cancers in female Canadians, the most rapidly increasing incidence is that of cervical cancer. The objective of this pilot study was to assess how HPV self-sampling might improve cervical cancer screening participation in both urban and rural settings in Saskatchewan, one of the most sparsely populated provinces in Canada. Study groups consisted of n = 250 participants to whom self-swabbing kits were mailed with instructions and n = 250 participants to whom kits were handed out in 6 urban and rural clinics. The inclusion criteria selected subjects aged 30-69 years who were Saskatchewan residents for at least 5 years with valid health coverage, had a cervix, and had no record of cervical cancer screening in 4 years. The returned samples were analyzed for specific HPV strains using the Roche Molecular Diagnostics Cobas 4800® System. The overall response rate was ~16%, with the response to the handout distribution being roughly double that of the mailout. While HPV positivity did not differ across the distribution groups, participants at a specific inner-city clinic reported significantly higher positivity to at least one HPV strain as compared to any other clinic and all mailouts combined. For this high-risk population, in-person handout of self-sampling kits may be the most effective means of improving screening.

Asunto(s)

Detección Precoz del Cáncer , Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Humanos , Femenino , Persona de Mediana Edad , Proyectos Piloto , Saskatchewan/epidemiología , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/virología , Adulto , Anciano , Detección Precoz del Cáncer/métodos , Infecciones por Papillomavirus/diagnóstico , Infecciones por Papillomavirus/virología , Papillomaviridae/genética , Papillomaviridae/aislamiento & purificación , Manejo de Especímenes/métodos , Tamizaje Masivo/métodos , Autocuidado

RESUMEN

Accurate specimen marking is crucial during breast cancer surgery to avoid misorientation, which can lead to inadequate re-excision and tumor recurrence. We studied the marking methods at various breast cancer centers to create a tool that would prevent specimen misorientation. An online questionnaire was used to survey marking procedures at major breast cancer centers in Hungary, and a tool was developed using a troubleshooting method. Twelve out of twenty units responded (60%). Nine use an institutionally standardized marking system. Less than half of the surgical teams found specimen mammograms to be unambiguous. In more than 70% of departments, pathologists were uncertain about breast specimen orientation. Ambiguous marking methods caused orientation errors in half of the cases, while unclear marking directions caused the rest. Most pathologists (85%) and surgeons (75%) believed that coronal plane specimen mammography would help solve the problem. A plastic specimen plate has been developed to anchor breast tissue to a coronal breast scheme as seen in mammography images, providing clear localization information throughout the surgical process. There is a lack of standardization in breast specimen orientation and marking in Hungary. An optimized orientation toolkit is being developed to ensure consistent interpretation of specimen mammograms by surgeons and pathologists.

Asunto(s)

Neoplasias de la Mama , Manejo de Especímenes , Humanos , Femenino , Neoplasias de la Mama/cirugía , Manejo de Especímenes/métodos , Mamografía/métodos , Encuestas y Cuestionarios , Hungría , Mama/cirugía , Mama/diagnóstico por imagen

RESUMEN

BACKGROUND: Oral microbiota comprises polymicrobial communities shaped by mutualistic coevolution with the host, contributing to homeostasis and regulating immune function. Nevertheless, dysbiosis of oral bacterial communities is associated with a number of clinical symptoms that ranges from infections to oral cancer. Peri-implant diseases are biofilm-associated inflammatory conditions affecting the soft and hard tissues around dental implants. Characterization and identification of the biofilm community are essential for the understanding of the pathophysiology of such diseases. For that sampling methods should be representative of the biofilm communities Therefore, there is a need to know the effect of different sampling strategies on the biofilm characterization by next generation sequencing. METHODS: With the aim of selecting an appropriate microbiome sampling procedure for periimplant biofilms, next generation sequencing was used for characterizing the bacterial communities obtained by three different sampling strategies two months after transepithelial abutment placement: adjacent periodontal crevicular fluid (ToCF), crevicular fluid from transepithelial abutment (TACF) and transepithelial abutment (TA). RESULTS: Significant differences in multiple alpha diversity indices were detected at both the OTU and the genus level between different sampling procedures. Differentially abundant taxa were detected between sample collection strategies, including peri-implant health and disease related taxa. At the community level significant differences were also detected between TACF and TA and also between TA and ToCF. Moreover, differential network properties and association patterns were identified. CONCLUSIONS: The selection of sample collection strategy can significantly affect the community composition and structure. TRIAL REGISTRATION: This research is part of a randomized clinical trial that was designed to assess the effect of transepithelial abutment surface on the biofilm formation. The trial was registered at Trial Registration ClinicalTrials.gov under the number NCT03554876.

Asunto(s)

Biopelículas , Implantes Dentales , Líquido del Surco Gingival , Microbiota , Humanos , Líquido del Surco Gingival/microbiología , Implantes Dentales/microbiología , Persona de Mediana Edad , Manejo de Especímenes/métodos , Femenino , Masculino , Secuenciación de Nucleótidos de Alto Rendimiento , Bacterias/clasificación , Bacterias/aislamiento & purificación , Anciano

RESUMEN

Volume electron microscopy encompasses a set of electron microscopy techniques that can be used to examine the ultrastructure of biological tissues and cells in three dimensions. Two block face techniques, focused ion beam scanning electron microscopy (FIB-SEM) and serial block face scanning electron microscopy (SBF-SEM) have often been used to study biological tissue samples. More recently, these techniques have been adapted to in vitro tissue culture samples. Here we describe step-by-step protocols for two sample embedding methods for in vitro tissue culture cells intended to be studied using SBF-SEM. The first focuses on cell pellet embedding and the second on en face embedding. En face embedding can be combined with light microscopy, and this CLEM workflow can be used to identify specific biological events by light microscopy, which can then be imaged using SBF-SEM. We systematically outline the steps necessary to fix, stain, embed and image adherent tissue culture cell monolayers by SBF-SEM. In addition to sample preparation, we discuss optimization of parameters for data collection. We highlight the challenges and key steps of sample preparation, and the consideration of imaging variables.

Asunto(s)

Microscopía Electrónica de Rastreo , Microscopía Electrónica de Rastreo/métodos , Animales , Humanos , Manejo de Especímenes/métodos , Adhesión del Tejido/métodos , Microscopía Electrónica de Volumen

RESUMEN

Introduction: In 2017, the Ministry of Health and Public Hygiene (MoH) of Burkina Faso designed and piloted a specimen transport system using the national courier services (La Poste BF) in 4 districts. Based on satisfactory performance indicators, the MoH set a vision aimed at scaling up this system to strengthen disease detection and surveillance of epidemic prone diseases across the country. This work describes the implementation process, performances, and lessons learned. Methodology: This work describes the implementation process, performances, and lessons learned. Under the leadership of the Directorate of Population Health Protection within the MoH, a stepwise approach was used to bring together multiple partners across sectors to develop the first needed documents including a guide, an implementation plan, Standard Operating Procedures, and data collection tools. Then, the execution phase included equipment purchase, trainings, and consensus on a financing mechanism. Key indicators were defined to allow performance monitoring. Result: The integrated biological specimen referral system (SITEB) was officially launched in January 2020 to transport human biological specimens of priority diseases including COVID-19 from district level to reference laboratories nationwide. As of December 31, 2022, La Poste BF transported 168,856 packages containing 206,314 specimens from all 13 regions. 99.66% of packages were delivered in <24 h as required, and 99.68% of specimens were in good condition at reception. COVID-19 specimens represented respectively 18% and 63% of samples transported in 2020 and 2021. Discussion: The political will combined with the experience gained during the pilot phase and the commitment and support from all stakeholders laid to the foundation of the effective implementation of this system. Collaboration between two government entities (MoH and Minister of Transport, Urban Mobility, and Road Safety) to benefit public health has led to reasonable pricing for sustainability. Although all documents integrate the "One Health" approach, the system ensures the transport of only human samples for now. Despite security constraints, Burkina Faso has successfully set up a system using the national postal service to ensure the routine transport of specimens for all diseases under laboratory surveillance including laboratory tests for HIV and TB from the district level to reference laboratories nationwide. This system has also proved to be useful and efficient in managing public health emergency.

Asunto(s)

COVID-19 , Burkina Faso , Humanos , Manejo de Especímenes , Derivación y Consulta , SARS-CoV-2

RESUMEN

BACKGROUND: Latinas experience the greatest cervical cancer incidence compared with other ethnic/racial groups in the United States (US) due in part to significant disparities in screening uptake. Social and structural conditions that impede access to and participation in screening include language barriers, concerns about documentation status, logistical issues (e.g., transportation, limited clinic hours), and cultural beliefs regarding modesty and promiscuity. To overcome these challenges, self-sampling for human papillomavirus (HPV) DNA testing has emerged as a potentially promising method for promoting cervical cancer screening among this population. Thus, this systematic review aimed to assess the acceptability of HPV self-sampling among US Latinas. METHODS: Using EBSCOhost and PubMed databases, we searched for studies published in the past two decades (2003-2023) that described participation in HPV self-sampling among Latinas. Eleven articles met inclusion criteria. RESULTS: The majority of studies were conducted in Florida, California, and Puerto Rico, were single-arm designs, and involved the use of community health workers and Spanish-language materials (e.g., brochures). Across studies, the majority of participants reported that self-sampling was acceptable with respect to ease of use, comfort (lack of pain), privacy, and convenience; however, some women were concerned about the accuracy of self-sampling or whether they had performed sample collection correctly. CONCLUSION: Given the high acceptability, self-collection of cervicovaginal samples for HPV testing may offer a feasible option for enhancing participation in cervical cancer screening in this population that encounters significant barriers to screening.

Asunto(s)

Detección Precoz del Cáncer , Hispánicos o Latinos , Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Humanos , Femenino , Infecciones por Papillomavirus/diagnóstico , Infecciones por Papillomavirus/virología , Infecciones por Papillomavirus/epidemiología , Estados Unidos/epidemiología , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/virología , Detección Precoz del Cáncer/métodos , Manejo de Especímenes/métodos , Aceptación de la Atención de Salud , Papillomaviridae/aislamiento & purificación , Papillomaviridae/genética , Adulto , Virus del Papiloma Humano