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Human papillomavirus (HPV) is the most prevalent sexually transmitted infection (STI) worldwide, with popular screening methods including the Papanicolaou test and HPV genotyping. However, in clinical practice, coinfections with other pathogens are often underestimated. Therefore, our study aims to describe the prevalence of STIs and vaginosis in urogenital samples from patients who had been tested exclusively for HPV genotyping. METHODS: This analytical, prospective, cross-sectional study included 408 males and females. Eligible participants had positive and negative HPV genotyping test results and agreed to early detection or had HPV antecedents. They provided the same urogenital samples used for HPV detection and, through our multiplex in-house PCR assay, we screened for Candida spp., Ureaplasma spp., Trichomonas vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, herpes simplex virus 1 and 2 (HSV), Mycoplasma spp., molluscum contagiosum virus (MCV), Treponema pallidum, Haemophilus spp., Staphylococcus aureus, and Klebsiella spp. The subsequent statistical analysis aimed to reveal correlations between HPV genotypes and the identified pathogens. RESULTS: Of the participants, 72.1% (n = 294) tested positive for HPV genotypes. HR-HPV (high-risk HPV) genotypes comprised 51 (8.1%), 66 (7.1%), and 58 (6.1%). Haemophilus spp., Ureaplasma spp., Candida spp., Staphylococcus aureus, and Mycoplasma spp. frequently co-occurred with HPV infection (p < 0.05). Gender-based variations were notorious for Ureaplasma spp., Mycoplasma spp., and MCV (p < 0.05). Coinfections were prevalent (43.9%), with a positive HPV result elevating the risk for Trichomonas vaginalis, Mycoplasma spp., Staphylococcus aureus, HSV, and MCV (OR > 1, p < 0.05). HPV 16 correlated with HSV and Ureaplasma spp., while HPV 6 was linked with HSV and MCV (p < 0.05). CONCLUSIONS: This screening strategy uncovered significant coinfections and associations between HPV genotypes and pathogens, underscoring the importance of routine screening to explore clinical implications in urogenital health.
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INTRODUCCIÓN: La diarrea aguda continúa siendo una de las principales causas de morbilidad en niños; sin embargo, el diagnóstico etiológico presenta limitaciones dada la baja sensibilidad de los métodos tradicionales. OBJETIVO: Describir los microorganismos identificados en niños que acudieron al Servicio de Urgencia (SU) de un hospital universitario en Santiago, Chile, por diarrea aguda y a los que se le solicitó panel molecular gastrointestinal. MÉTODOS: Se revisaron fichas clínicas y resultados de panel gastrointestinal realizados entre junio de 2017 y marzo de 2020. RESULTADOS: Se incluyeron 198 pacientes, edad promedio de 54,5 meses y 60,6% (120/198) de sexo masculino. La positividad del panel fue de 78,8% (156/198) con 35,3% (55/156) de las muestras polimicrobianas. Se identificaron 229 microorganismos, de los cuales 72,9% (167/229) corresponden a bacterias, 25,8% (59/229) a virus y 1,3% (3/229) a parásitos. Destacaron Campylobacter spp. y Escherichia coli enteropatógena (ECEP) como las bacterias más frecuentemente identificadas. Los pacientes con detección de Campylobacter spp. presentaron con mayor frecuencia fiebre (p = 0,00). ECEP se aisló principalmente (82,5%) en muestras polimicrobianas. DISCUSIÓN: Los resultados enfatizan el potencial que poseen los estudios moleculares para mejorar el diagnóstico etiológico de la diarrea, pero a la vez llevan a cuestionar el rol patogénico de algunos microorganismos identificados.
BACKGROUND: Acute diarrhea continues to be one of the main causes of morbidity in children, however the etiologica diagnosis presents limitations given the low sensitivity of traditional methods. AIM: To describe the microorganisms identified in children who attended the emergency department (ED) in Santiago, Chile, due to acute diarrhea and to whom a gastrointestinal panel was requested as part of their study. MATERIAL AND METHODS: Clinical records and results of the gastrointestinal panel carried out between June 2017 and March 2020 were reviewed. RESULTS: 198 patients were included, the average age was 54.5 months and 60.6% (120/198) were males. Positivity was 78.8% (156/198) with 35.3% (55/156) of the samples being polymicrobial. 229 microorganisms were identified, of which 72.9% (167/229) corresponded to bacteria, 25.8% (59/229) to viruses, and 1.3% (3/229) to parasites. Campylobacter spp. and enteropathogenic Escherichia coli (EPEC) were the most frequently identified bacteria. Patients with detection of Campylobacter spp. presented a higher frequency of fever (p = 0.00). EPEC was isolated in 82.5% of the cases in polymicrobial samples. DISCUSSION: The results emphasize the potential of molecular studies to improve the etiological diagnosis of diarrhea and at the same time lead to question the pathogenic role of some microorganisms.
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Humanos , Masculino , Feminino , Diarreia/diagnóstico , Fezes/microbiologia , Parasitos/isolamento & purificação , Estações do Ano , Bactérias/isolamento & purificação , Vírus/isolamento & purificação , Chile , Estudos Retrospectivos , Diarreia/etiologia , Diarreia/epidemiologia , Serviço Hospitalar de Emergência , Fezes/parasitologiaRESUMO
A multiplex PCR method was developed for the simultaneous detection of murine norovirus (MNV-1) as a surrogate for human norovirus (HuNoV) GI and GII, Salmonella spp., Shigella spp., and Shiga toxin producing Escherichia coli (STEC) in fresh produce. The toxicity of the glycine buffer on bacterial pathogens viability was evaluated. The growth of each of the three pathogens (previously stressed) was evaluated at 35 and 41.5 °C in modified buffered peptone water (mBPW) and trypticase soy broth (TSB), supplemented with vancomycin, novobiocin and brilliant green at two concentration levels. The selected conditions for simultaneous enrichment were: 41.5 °C/mBPW/supplemented with 8 ppm vancomycin, 0.6 ppm novobiocin and 0.2 ppm brilliant green. The pathogens and aerobic plate count (APC) growth was evaluated in the enrichment of lettuce, coriander, strawberry and blackberry under the best enrichment conditions. Starting from 1 to 10 CFU/mL, Salmonella reached from 7.63 to 8.91, Shigella 6.81 to 7.76 and STEC 7.43 to 9.27 log CFU/mL. The population reached for the APC was 5.11-6.56 log CFU/mL. Simultaneous detection by PCR was done using designed primers targeting invA, ipaH, stx1 and stx2 genes, and MNV-1. The detection sensitivity was 10-100 PFU for the MNV-1 and 1-10 CFU for each pathogenic bacteria. This protocol takes 6 h for MNV-1 and 24 h for Salmonella spp., Shigella spp., and STEC detection from the same food portion. In total, 200 samples were analyzed from retail markets from Queretaro, Mexico. Two strawberry samples were positive for HuNoV GI and one lettuce sample was positive for STEC. In conclusion, the method developed in this study is capable of detecting HuNoV GI and GII, Salmonella spp., Shigella spp and STEC from the same fresh produce sample.
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Coriandrum , Contaminação de Alimentos/análise , Microbiologia de Alimentos/métodos , Fragaria , Lactuca , Rubus , Coriandrum/microbiologia , Coriandrum/virologia , Fragaria/microbiologia , Fragaria/virologia , Frutas/microbiologia , Frutas/virologia , Lactuca/microbiologia , Lactuca/virologia , Reação em Cadeia da Polimerase Multiplex , Norovirus/isolamento & purificação , Novobiocina , Rubus/microbiologia , Rubus/virologia , Salmonella/isolamento & purificação , Escherichia coli Shiga Toxigênica/isolamento & purificação , Shigella/isolamento & purificação , VancomicinaRESUMO
INTRODUCTION: Epstein-Barr Virus (EBV) infection prevails in underdeveloped and developing countries. The tonsils seem to be candidate replication sites for EBV and some studies have exposed a close association among viral infections and chronic tonsillitis. The objective of this study was identifying the EBV prevalence in Mexican patients who had undergone tonsillectomy because of chronic tonsillitis. METHODOLOGY: Frozen tissues and medical records were obtained from 50 Mexican patients. DNA was extracted and subjected to PCR to amplify the EBER-2 region of EBV. Next, the patients were classified according to general and clinical characteristics searching a relation with the EBV-DNA positivity. RESULTS: EBV genome was detected in 46% (23/50) of the analysed tonsil tissues. Trends were found regarding the relationship of viral presence with lower values in terms of age (6.1 ± 2.8 vs 7.6 ± 3.7) , a greater degree of hypertrophy (3.5 ± 0.4 vs 3.0 ± 0.6) and an increase in the number of episodes of tonsillitis (11 ± 7.4 vs 9 ± 6.5). CONCLUSIONS: The prevalence found of EBV-DNA positivity in tonsillar tissues from patients diagnosed with chronic tonsillitis , supports the fact that palatine tonsils can be occupied by EBV and highlights the importance of conducting future studies focused on understanding the role of the EBV infection in chronic inflammatory processes in the population involved in this study.
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DNA Viral/análise , Infecções por Vírus Epstein-Barr/epidemiologia , Infecções por Vírus Epstein-Barr/virologia , Herpesvirus Humano 4/isolamento & purificação , Tonsila Palatina/virologia , Tonsilite/epidemiologia , Tonsilite/virologia , Adolescente , Criança , Pré-Escolar , Doença Crônica , DNA Viral/genética , Feminino , Herpesvirus Humano 4/genética , Humanos , Lactente , Recém-Nascido , Masculino , México/epidemiologia , Reação em Cadeia da Polimerase , PrevalênciaRESUMO
The present study was carried out to determine if group 16SrIV phytoplasmas, causing lethal yellowing (LY) disease, are present in Haplaxius crudus Van Duzee (Hemiptera: Cixiidae) insects associated with palms in Yucatán, Mexico. Haplaxius crudus feral insects were captured from palm foliage at two locations (Chicxulub Puerto and CICY, Mérida, where LY-type diseases are active) and evaluated individually for the presence of phytoplasma DNA by a group 16SrIV-specific nested PCR assay. The results showed positive detection in H. crudus insects in a proportion of 2.7% (of the total 2726 analyzed) during a 3-year period of study. The percentage of detection was different for each site, 5.9% positive of 799 insects from Mérida and 1.7% of 1927 from Chicxulub Puerto. Positive detections were also obtained in extracts from 5.3 to 1.2% of males and females, respectively. Sequencing and in silico RFLP and phylogenetic analyses of PCR-amplified rDNA products indicated that H. crudus insects from Chicxulub Puerto harbored phytoplasma strains of subgroups 16SrIV-A or 16SrIV-D, whereas in insects from Mérida the strains found were 16SrIV-A, 16SrIV-D or 16SrIV-E. The diversity of subgroup strains detected in H. crudus coincided with strains previously identified in palms showing LY-type disease syndromes in Yucatán thereby implicating H. crudus as a candidate vector of 16SrIV phytoplasmas in this region of Mexico.
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Abstract: INTRODUCTION: Before 2004, the occurrence of acute Chagas disease (ACD) by oral transmission associated with food was scarcely known or investigated. Originally sporadic and circumstantial, ACD occurrences have now become frequent in the Amazon region, with recently related outbreaks spreading to several Brazilian states. These cases are associated with the consumption of açai juice by waste reservoir animals or insect vectors infected with Trypanosoma cruzi in endemic areas. Although guidelines for processing the fruit to minimize contamination through microorganisms and parasites exist, açai-based products must be assessed for quality, for which the demand for appropriate methodologies must be met. METHODS: Dilutions ranging from 5 to 1,000 T. cruzi CL Brener cells were mixed with 2mL of acai juice. Four Extraction of T. cruzi DNA methods were used on the fruit, and the cetyltrimethyl ammonium bromide (CTAB) method was selected according to JRC, 2005. RESULTS: DNA extraction by the CTAB method yielded satisfactory results with regard to purity and concentration for use in PCR. Overall, the methods employed proved that not only extraction efficiency but also high sensitivity in amplification was important. CONCLUSIONS: The method for T. cruzi detection in food is a powerful tool in the epidemiological investigation of outbreaks as it turns epidemiological evidence into supporting data that serve to confirm T. cruzi infection in the foods. It also facilitates food quality control and assessment of good manufacturing practices involving acai-based products.
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Humanos , Animais , Trypanosoma cruzi/isolamento & purificação , Contaminação de Alimentos , DNA de Protozoário/isolamento & purificação , Parasitologia de Alimentos , Doença de Chagas/transmissão , Euterpe/parasitologia , Trypanosoma cruzi/genética , Reação em Cadeia da Polimerase , Surtos de Doenças , Doença de Chagas/epidemiologiaRESUMO
The recently described trypanosome Lotmaria passim is currently considered the most predominant trypanosomatid in honey bees worldwide and could be a factor in honey bee declines. For a specific and quick detection of this pathogen, we developed primers based on the SSU rRNA and gGAPDH genes for the detection of L. passim in Chilean honey beehives. PCR products amplified and sequenced for these primers shared 99-100% identity with other sequences of L. passim. The designed primers were specific and we were able to detect a high prevalence (40-90%) of L. passim in bee hives distributed throughout Chile. Our described PCR-based method offers a feasible and specific detection of L. passim in any honey bee samples.
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Abelhas/parasitologia , Trypanosomatina/genética , Animais , Chile , Primers do DNA , DNA de Protozoário/química , Filogenia , Reação em Cadeia da Polimerase/métodos , Trypanosomatina/isolamento & purificaçãoRESUMO
Bartonellosis and rickettsiosis are commonly reported in Peru. In order to detect Bartonella sp. and Rickettsia sp. in fleas, ticks and lice, specimens from five distinct locations in Peru (Marizagua, Cajaruro, Jamalca, Lonya Grande and El Milagro) were collected and screened for the presence of these bacteria using PCR and later confirmation by DNA sequencing. The specimens collected were distributed in 102 pools (76 Ctenocephalides felis, 2 Ctenocephalides canis, 16Pulex irritans, 5 Pediculus humanus, 2 Rhiphicephalus sanguineus, and 1 Boophilus spp.), whereBartonella was detected in 17 pools (6 of C. felis, 9 of P. irritans, 1 of C. canis, and 1 P. humanus). Also, Rickettsia was detected in 76 pools (62 C. felis, 10 P. irritans, 2 P. humanus, and 2 C. canis). Bartonella clarridgeiae was detected in C. felis, C. canis and P. irritans pools at 5.3%, 50% and 12.5%, respectively. Bartonella rochalimae was detected in one C. felis and two P. irritans pools at 1.3% and 12.5%, respectively. Furthermore, B. henselae was detected in one C. felis pool and one P. humanus pool corresponding to 1.3% and 20%, respectively; andBartonella spp. was also found in 5 pools of P. irritans at 31.3%. Additionally, R. felis was detected in C. felis, C. canis and P. irritans pools at 76.3%, 100% and 37.5%, respectively; and Rickettsia spp. was detected in C. felis, P. irritans and P. humanus pools at 5.3%, 25% and 40%, respectively. These results demonstrate the circulation of these bacteria in Peru
La Bartonellosis y la Rickettsiosis son enfermedades comúnmente reportadas en Perú. Con el propósito de detectar Bartonella sp. y Rickettsia sp. especímenes de pulgas, garrapatas y piojos de cinco localidades del Perú (Marizagua, Cajaruro, Jamalca, Lonya Grande and El Milagro) fueron colectadas y analizadas. Para la detección se usó PCR y una posterior confirmación con secuenciamiento de DNA. Los especímenes colectados fueron agrupados en 102 pools (76 Ctenocephalides felis, dos Ctenocephalides canis, 16Pulex irritans, cinco Pediculus humanus, dos Rhiphicephalus sanguineus, y un Boophilus spp.). Bartonella fue detectada en 17 pools (seis de C. felis, nueve de P. irritans, uno de C. canis, y uno de P. humanus). Rickettsia fue detectada en 76 pools (62 de C. felis, 10 de P. irritans, dos de P. humanus, y dos de C. canis). Bartonella clarridgeiae fue detectada en C. felis (5.3% especímenes), C. canis (50%) y P. irritans (12.5%). Bartonella rochalimae fue detectada en C. felis (1.3%) y P. irritans (12.5%). Además, se detectó B. henselae en C. felis (1.3%) y P. humanus (20%). Bartonella spp. también se encontró en P. irritans (31,3%). Además, se detectó R. felis en C. felis (76.3%), C. canis (100%) y P. irritans (37.5%), y Rickettsia spp. se detectó en C. felis (5,3%), P. irritans (25%) y P. humanus (40%). Estos resultados demuestran la circulación de estas bacterias en el Perú
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Ochratoxin A (OTA) is one of the most abundant food-contaminating mycotoxins. Its occurrence in several agricultural commodities has been considered a worldwide problem. This toxin is mainly produced by Aspergillus species. OTA has nephrotoxic, immunosuppressive, and carcinogenic effects and consequently the contamination with this toxin represents a high risk for human and animal health. In the last 5 years, several investigators have applied molecular methods in order to develop PCR assays for identifying and quantifying OTA-producing fungi in coffee beans samples. The main objective is to allow the detection of microorganisms capable of producing OTA, preferentially prior to ochratoxin production and accumulation. In this contribution several of these attempts will be reviewed and discussed.
Dentre as micotoxinas que contaminam produtos destinados à alimentação humana e animal, a ocratoxina A (OTA) é uma das mais freqüentemente encontrada. A sua ocorrência em vários produtos agrícolas tem sido considerada um problema de amplitude mundial. Esta toxina é produzida principalmente por fungos do gênero Aspergillus. A OTA tem efeitos nefrotóxico, imunossupressor e carcinogênico. A contaminação de alimentos com esta toxina representa risco para a saúde animal e humana. Nos últimos cinco anos, vários investigadores têm desenvolvido métodos moleculares para identificação e quantificação de fungos produtores de OTA em amostras de grãos de café. O objetivo desta revisão é apresentar e discutir as várias estratégias desenvolvidas recentemente para a detecção de fungos potencialmente produtores de OTA.
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This study has detected BPV-1 DNA sequences in wart, blood and plasma samples collected from animals affected by papillomatosis, suggesting viral presence inside the cell. We sellected an animal in which we could detect BPV-1 DNA sequences in wart, blood, placenta and amniotic liquid samples and her offspring which presented BPV-1 DNA sequences in blood sample collected immediately after birth. These results show a possible vertical transmission of BPV-1.
Neste estudo detectou-se DNA de BPV-1 em: verruga, sangue e plasma de animais afetados por papilomatose. Estes resultados trazem evidências sobre possível localização intracelular do BPV-1 no sangue. Avaliamos um animal afetado por papilomatose e positivo para BPV-1 em amostras de verruga, sangue, placenta e líquido amniótico e que teve sua cria recém-nascida também positiva para BPV-1 no sangue. Estes resultados indicam possível transmissão vertical do BPV-1.
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This study has detected BPV-1 DNA sequences in wart, blood and plasma samples collected from animals affected by papillomatosis, suggesting viral presence inside the cell. We sellected an animal in which we could detect BPV-1 DNA sequences in wart, blood, placenta and amniotic liquid samples and her offspring which presented BPV-1 DNA sequences in blood sample collected immediately after birth. These results show a possible vertical transmission of BPV-1.
Neste estudo detectou-se DNA de BPV-1 em: verruga, sangue e plasma de animais afetados por papilomatose. Estes resultados trazem evidências sobre possível localização intracelular do BPV-1 no sangue. Avaliamos um animal afetado por papilomatose e positivo para BPV-1 em amostras de verruga, sangue, placenta e líquido amniótico e que teve sua cria recém-nascida também positiva para BPV-1 no sangue. Estes resultados indicam possível transmissão vertical do BPV-1.
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This study has detected BPV-1 DNA sequences in wart, blood and plasma samples collected from animals affected by papillomatosis, suggesting viral presence inside the cell. We sellected an animal in which we could detect BPV-1 DNA sequences in wart, blood, placenta and amniotic liquid samples and her offspring which presented BPV-1 DNA sequences in blood sample collected immediately after birth. These results show a possible vertical transmission of BPV-1.
Neste estudo detectou-se DNA de BPV-1 em: verruga, sangue e plasma de animais afetados por papilomatose. Estes resultados trazem evidências sobre possível localização intracelular do BPV-1 no sangue. Avaliamos um animal afetado por papilomatose e positivo para BPV-1 em amostras de verruga, sangue, placenta e líquido amniótico e que teve sua cria recém-nascida também positiva para BPV-1 no sangue. Estes resultados indicam possível transmissão vertical do BPV-1.