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To evaluate the effectiveness of a home exercise program called Home Exercise Booklet for People Living with Human T Lymphotropic Virus 1 (HTLV-1). This is a methodological study of content validation with expert judges. A questionnaire with a Likert scale was applied, containing 16 items referring to the content domain. Descriptive statistics were used to obtain the content validity index. In total, 46 judges participated, 24 physiotherapists (PG) and 22 professionals from other health areas specializing in methodological studies and HTLV-1 (EG). In the validation process, each evaluator judged the technology and scored their considerations. In the end, we obtained the following results for the Content Validity Index (CVI): PG CVI: 94.3%, GE CVI: 93.4%. Although the index was sufficient to consider the technology validated, modifications were made to the second and final version of the booklet, considering the judges' observations and suggestions, which we consider relevant. The technology proved to be valid for use with the target audience. The development and validation of this product provides support to help prevent functional decline in people living with HTLV-1; standardize guidelines for physiotherapy professionals who monitor these issues; start a home exercise program aimed at other comorbidities; open the possibility of creating and validating home exercise programs with other comorbidities.
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Terapia por Ejercicio , Virus Linfotrópico T Tipo 1 Humano , Humanos , Terapia por Ejercicio/métodos , Encuestas y Cuestionarios , Femenino , Infecciones por HTLV-I/prevención & control , Masculino , Adulto , Personas con Discapacidad/rehabilitación , Persona de Mediana Edad , Ejercicio FísicoRESUMEN
BACKGROUND: Approximately 80% of infected women infected by Chlamydia trachomatis are asymptomatic, although this infection can lead to serious complications in the female reproductive tract. Few data on Chlamydia infection and genotypes are available in Amazonian communities. OBJECTIVES: To describe the prevalence of and associated factors and to identify the genotypes of sexual C. trachomatis infection in female university students in different urban centers (capital and interiors) in the Brazilian state of Pará, in the eastern Amazon region. METHODS: A cross-sectional study was performed among young women attending public universities in four different urban centers in the eastern Amazon region. They were invited to participate in the studt and cervical secretions were collected for molecular diagnosis of C. trachomatis. We utilized amplification of the ompA gene by nested PCR. Positive samples were genotyped by nucleotide sequencing. Study participants completed a questionnaire on social, epidemiological, and reproductive health variables. A Qui-square and Binominal regression test were used to evaluate the degree of association of these variables with the infection. RESULTS: A total of 686 female students was included in the study. The overall prevalence of C. trachomatis was 11.2% (77/686). The prevalence of this infection was higher in interiors (15.2% vs 9.5%/ p: 0.0443). Female university students who do not have a sexual partner (11.8%/p <0.008), who do not use a condom in their sexual relations (17.8%/p <0.0001) and who reported having suffered a miscarriage (32%/p <0.0001) have high chances of acquiring this sexual infection. The ompA gene was sequenced in only 33 (42.8%) samples, revealing the genotype J was the most frequent (27.2% [9/33]), followed by genotypes D (24.2% [8/33]), and then genotypes F (18.2% [6/33]), E (15.1% [5/33]) K (6.1% [2/33]), Ia (6.1% [2/33]), and G (3.1% [1/33]). CONCLUSIONS: The high prevalence of sexual infection by C. trachomatis in the female university students from the interior of the state of Pará, individuals with no fixed sexual partner, those that had had a miscarriage, the students that do not use condoms in their sexual relations. The genotype J of C. trachomatis genotypes was the most frequent. These data are important to help defining the epidemiological effects of chlamydial infections in Amazonian populations.
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Aborto Espontáneo , Infecciones por Chlamydia , Embarazo , Humanos , Femenino , Chlamydia trachomatis/genética , Universidades , Prevalencia , Ciudades/epidemiología , Brasil/epidemiología , Estudios Transversales , Infecciones por Chlamydia/epidemiología , Infecciones por Chlamydia/diagnóstico , GenotipoRESUMEN
Introduction: After three years since the beginning of the pandemic, the new coronavirus continues to raise several questions regarding its infectious process and host response. Several mutations occurred in different regions of the SARS-CoV-2 genome, such as in the spike gene, causing the emergence of variants of concern and interest (VOCs and VOIs), of which some present higher transmissibility and virulence, especially among patients with previous comorbidities. It is essential to understand its spread dynamics to prevent and control new biological threats that may occur in the future. In this population_based retrospective observational study, we generated data and used public databases to understand SARS-CoV-2 dynamics. Methods: We sequenced 1,003 SARS-CoV-2 genomes from naso-oropharyngeal swabs and saliva samples from Pará from May 2020 to October 2022. To gather epidemiological data from Brazil and the world, we used FIOCRUZ and GISAID databases. Results: Regarding our samples, 496 (49.45%) were derived from female participants and 507 (50.55%) from male participants, and the average age was 43 years old. The Gamma variant presented the highest number of cases, with 290 (28.91%) cases, followed by delta with 53 (5.28%). Moreover, we found seven (0.69%) Omicron cases and 651 (64.9%) non-VOC cases. A significant association was observed between sex and the clinical condition (female, p = 8.65e-08; male, p = 0.008961) and age (p = 3.6e-10). Discussion: Although gamma had been officially identified only in December 2020/January 2021, we identified a gamma case from Belém (capital of Pará State) dated May 2020 and three other cases in October 2020. This indicates that this variant was circulating in the North region of Brazil several months before its formal identification and that Gamma demonstrated its actual transmission capacity only at the end of 2020. Furthermore, the public data analysis showed that SARS-CoV-2 dispersion dynamics differed in Brazil as Gamma played an important role here, while most other countries reported a new infection caused by the Delta variant. The genetic and epidemiological information of this study reinforces the relevance of having a robust genomic surveillance service that allows better management of the pandemic and that provides efficient solutions to possible new disease-causing agents.
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COVID-19 , SARS-CoV-2 , Humanos , Femenino , Masculino , Adulto , SARS-CoV-2/genética , Brasil/epidemiología , COVID-19/epidemiología , Análisis de DatosRESUMEN
The human T-lymphotropic virus type 1 (HTLV-1), isolated in 1980, causes T-cell leukemia/lymphoma in adulthood, a type of lymphoproliferative disease, and chronic HTLV-1-associated myelopathy, a disease that causes paralysis of the lower limbs, which occur in about 5% of cases in this viral infection. This study aimed to establish the hematological profile of patients with HTLV-1 infection in Belém do Pará, describing the hematological parameters under study, estimating the frequency of lymphocytic atypical, and associating the hematological profile with diseases and symptoms. Hematologic data from 202 individuals were analyzed, including 87 HTLV-1 infected individuals and 115 non-HTLV-1 infected individuals as a control group, composed, at a great part, of relatives of the infected. The seroprevalence of HTLV-1 infection was observed in 71.3% of female individuals, with predominance in the group older than 50 years (44.8%). The analysis of hematological parameters showed a significant difference in the counts of the segmented cells (p = 0.0303) and eosinophils (p = 0.0092) in HTLV-1 carriers. Lymphocytic atypical was a finding present only in HTLV-1 carriers (p = 0.0001). There was no high frequency in the leukocyte counts of those infected by HTLV-1 not among them concerning a significant increase or decrease. It is concluded that HTLV-1 infection is prominent in women over 50 years old. The hematological profile of those infected shows a reduction of segmented cells, an increase of eosinophils, and the presence of atypical lymphocytes. The hematological profile of the HTLV-1 carrier should always be evaluated to identify early some diseases associated with the infection.
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BACKGROUND: The Human Papillomavirus (HPV) and Chlamydia trachomatis are the most prevalent Sexually Transmitted Infections (STIs) worldwide, and are associated cervical cancer and pelvic inflammatory disease, respectively. However, 80% of women testing positive are asymptomatic. In the Amazon region, young women, in particular, are widely exposed to the infections and their consequences. OBJECTIVES: Determine the prevalence of sexual infection by HPV and C. trachomatis in young, sexually-active women treated at a university health program in a large city of the Brazilian Amazon region. METHODS: We amplified the L1 gene of HPV. We amplified ompA gene of C. trachomatis by nested PCR, and the study participants filled in a questionnaire on their social, epidemiological, and reproductive health characteristics. The data were analyzed using the Odds Ratio, to evaluate the degree of association of these variables with the observed infections. RESULTS: The prevalence of infection by HPV was 15.5% (47/303). This infection was recorded in 32.2% of the women of less than 25 years of age (OR:3.02 [CI95%] = 1.32-6.92; p = 0.014), 17.9% of the single women (OR: 2.41 [CI95%] = 1.22-4.75; p = 0.014), 23.8% of the women that reported having first sexual intercourse at less than 15 years of age (OR: 2.22 [CI95%] = 1.16-4.23; p = 0.021), 20% of those that reported having had more than one sexual partner during their lifetime (OR: 3.83 [CI95%] = 1.56-9.37; p = 0.003), and in 28.3% that use oral contraceptives (CI95% = 1.33-5.43; p = 0.008). The prevalence of sexual infection by C. trachomatis was 4.6% (14/303), and this bacterium was present in 16.1% of the young women of less than 25 years of age (OR: 2.86 [CI95%] = 1.33-5.43; p = 0.008). CONCLUSIONS: We found a high prevalence of HPV in young, unmarried women who started their sex lives early, who had several sexual partners in their lives and who used oral contraceptives. The prevalence of C. trachomatis was high only in young women. Our data are in accordance with other studies in Brazil and in the world and may serve to base the formulation of diagnostic and screening measures for these infections in women in the Amazon.
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Alphapapillomavirus , Infecciones por Chlamydia , Infecciones por Papillomavirus , Brasil/epidemiología , Infecciones por Chlamydia/epidemiología , Chlamydia trachomatis , Anticonceptivos Orales , Femenino , Humanos , Papillomaviridae/genética , Infecciones por Papillomavirus/epidemiología , Prevalencia , Factores de Riesgo , Conducta SexualRESUMEN
BACKGROUND: Human T-lymphotropic virus 1 (HTLV-1) is etiologically associated with the chronic inflammatory neurodegenerative disease HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) Annexin A1 (AnxA1) is an anti-inflammatory protein with proposed neuroprotective and anti-neuroinflammatory functions. We hypothesized that ANXA1 gene expression may be dysregulated in HTLV-1-infected HAM/TSP patients. METHODS: This study involved 37 individuals infected with HTLV-1, including 21 asymptomatic (AS) carriers and 16 with HAM/TSP, and a control group of 30 individuals negative for HTLV-1 and HTLV-2. For AS HTLV-1-positive and HAM/TSP patients, ANXA1 and formyl peptide receptor (FPR1, FPR2 and FPR3) expression and HTLV-1 proviral load (PVL) in peripheral blood cells were evaluated by real-time quantitative PCR (qPCR), and plasma AnxA1 levels were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: ANXA1 gene expression was increased in the AS group compared with the HAM/TSP and control groups, but the differences were not statistically significant. FPR1 gene expression was higher in patients with HTLV-1 than in controls (AS, p = 0.0032; HAM/TSP, p < 0.0001). Plasma AnxA1 levels were higher in the AS group than in the HAM/TSP group (p = 0.0045), and PVL was higher in patients with HAM/TSP than in AS individuals (p = 0.0162). The use of a combined ROC curve using Annexin 1 levels and proviral load significantly increased the sensitivity and specificity to predict progression to HAM/TSP (AUC = 0.851 and AUC = 0.937, respectively, to AUC = 1000). CONCLUSIONS: Our results suggest that AnxA1 may be dysregulated in HAM/TSP patients. Serological detection of AnxA1 in association with proviral load may provide a prognostic biomarker for HTLV-1-associated neurodegenerative disease.
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Anexina A1/sangre , Virus Linfotrópico T Tipo 1 Humano/aislamiento & purificación , Paraparesia Espástica Tropical/diagnóstico , Adulto , Anexina A1/genética , Biomarcadores/sangre , Progresión de la Enfermedad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Paraparesia Espástica Tropical/sangre , Paraparesia Espástica Tropical/virología , Pronóstico , Curva ROC , Sensibilidad y Especificidad , Carga ViralRESUMEN
BACKGROUND: The forkhead box protein 3 (FOXP3) transcription factor is one of the main markers of immunological suppression in different pathological profiles, and the presence of polymorphic variants may alter the gene expression of this factor. Despite descriptions of an association between the presence of the rs2232365 polymorphism and chronic diseases, the role of the sex variant in this context has not yet been elucidated, as the FOXP3 gene is located on the human sex chromosome X. RESULTS: To contribute to this topic, 323 women and 373 men were enrolled in the study, of which 101 were diagnosed with chronic viral liver diseases (39 women and 62 men), 67 with HTLV-1 infection (44 women and 23 men), 230 with coronary artery disease (91 women and 139 men) and 298 healthy and uninfected blood donors (149 women and men). They were genotyped for the rs2232365 polymorphism. The rs2232365 polymorphism was associated with clinical and pathological aspects and biomarkers of viral infections only in men, with functional differences between different infections. CONCLUSIONS: A relationship is suggested between sex and FOXP3 rs2232365 polymorphism, resulting in different biological repercussions.
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Factores de Transcripción Forkhead/metabolismo , Genotipo , Infecciones por HTLV-I/genética , Virus Linfotrópico T Tipo 1 Humano/fisiología , Factores Sexuales , Adulto , Estudios de Casos y Controles , Enfermedad Crónica , Enfermedad de la Arteria Coronaria/genética , Femenino , Factores de Transcripción Forkhead/genética , Predisposición Genética a la Enfermedad , Infecciones por HTLV-I/inmunología , Humanos , Masculino , Polimorfismo de Nucleótido SimpleRESUMEN
SAMHD1, a host dNTPase, acts as a retroviral restriction factor by degrading the pool of nucleotides available for the initial reverse transcription of retroviruses, including HTLV-1. Polymorphisms in the SAMDH1 gene may alter the enzymatic expression and influence the course of infection by the virus. The present study investigated the effect of polymorphisms on HTLV-1 infection susceptibility and on progression to disease in 108 individuals infected by HTLV-1 (47 symptomatic and 61 asymptomatic) and 100 individuals in a control group. SAMHD1 rs6029941 (G/A) genotyping and HTLV-1 proviral load measurements were performed using real-time PCR and plasma IFN-α was measured by ELISA. Polymorphism frequency was not associated with HTLV-1 infection susceptibility or with the presence of symptoms. The proviral load was significantly higher in symptomatic individuals with the G allele (p = 0.0143), which presented lower levels of IFN-α (p = 0.0383). SAMHD1 polymorphism is associated with increased proviral load and reduced levels of IFN-α in symptomatic patients, and may be a factor that contributes to the appearance of disease symptoms.
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Infecciones por HTLV-I , Virus Linfotrópico T Tipo 1 Humano , Proteína 1 que Contiene Dominios SAM y HD , Carga Viral , Infecciones por HTLV-I/genética , Humanos , Provirus , Proteína 1 que Contiene Dominios SAM y HD/genéticaRESUMEN
BACKGROUND: Chlamydia trachomatis is the most prevalent bacterial sexually transmitted infection (STI) in the world. Approximately 80% of infected women are asymptomatic, although this infection can lead to serious complications in the female reproductive tract. Few data on Chlamydia infection are available in rural Amazonian communities. OBJECTIVES: To evaluate the prevalence of sexual C. trachomatis infection in women from Marajó Archipelago communities in the Amazon region of Brazil and to identify associated factors and genotypes. METHODS: We utilized amplification of the ompA gene by nested PCR. Positive samples were genotyped by sequencing. Study participants completed a questionnaire on social, epidemiological, and reproductive health variables. A Poisson regression was used to evaluate the degree of association of these variables with the infection. RESULTS: The sexual infection by C. trachomatis was observed in 4% (16/393) of the subjects, and was more often found in women aged ≤25 (14.3%; 95% CI = 2.83-35.47; p <0.001), and in women with a household income of less than one Brazilian monthly minimum wage (5.2%; 95% CI = 1.33-11.37; p = 0.014). The ompA gene was sequenced in 13 samples, revealing F genotypes (38.4%, n = 5), D (23%, n = 3), E (15.3%, n = 2), Ia (7.6%, N = 1), J (7.6%, n = 1) and B (7.6%, n = 1). CONCLUSIONS: We recorded a high prevalence of sexual infection by C. trachomatis in young and poor women from the interior of the Brazilian Amazon. This high prevalence and the frequencies of the main genotypes were similar to those found in major Brazilian urban centers. Our results reinforce the importance of the screening of this neglected infection, and the prevention of later sequelae in young women from rural and urban areas of Brazil.
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Infecciones por Chlamydia/epidemiología , Chlamydia trachomatis/fisiología , Islas/epidemiología , Adolescente , Adulto , Anciano , Brasil/epidemiología , Chlamydia trachomatis/genética , Femenino , Humanos , Persona de Mediana Edad , Prevalencia , Salud Reproductiva , Factores de Riesgo , Conducta Sexual , Adulto JovenRESUMEN
The Human T-cell Lymphotropic Virus (HTLV-1) is a Deltaretrovírus that was first isolated in the 1970s, and associated with Adult T-cell Leucemia-Lymphoma (ATLL), and subsequently to Tropical Spastic Paraparesis-Myelopathy (TSP/HAM). The genetic diversity of the virus varies among geographic regions, although its mutation rate is very low (approximately 1% per thousand years) in comparison with other viruses. The present study determined the genetic diversity of HTLV-1 in the metropolitan region of Belém, in northern Brazil. Blood samples were obtained from patients at the UFPA Tropical Medicine Nucleus between January 2010 and December 2013. The DNA was extracted and the PX region of the HTLV was amplified using nested PCR. The positive samples were then digested using the Taq1 enzyme for the identification and differentiation of the HTLV-1 and HTLV-2. The 5'LTR region of the positive HTLV-1 samples were amplified by nested PCR, and then sequenced genetically. The phylogenetic analysis of the samples was based on the maximum likelihood method and the evolutionary profile was analyzed by the Bayesian approach. Overall, 78 samples tested positive for HTLV-1, and 44 were analyzed here. The aA (cosmopolitan-transcontinental) subtype was recorded in all the samples. The following evolutionary rates were recorded for the different subtypes-a: 2.10-3, b: 2.69. 10-2, c: 6.23. 10-2, d: 3.08. 10-2, e: 6. 10-2, f: 1.78. 10-3, g: 2.2. 10-2 mutations per site per year. The positive HTLV-1 samples tested in the present study were characterized by their low genetic diversity and high degree of stability.
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Variación Genética , Virus Linfotrópico T Tipo 1 Humano/genética , Leucemia-Linfoma de Células T del Adulto/genética , Secuencias Repetidas Terminales , Adulto , Brasil/epidemiología , Femenino , Humanos , Leucemia-Linfoma de Células T del Adulto/enzimología , MasculinoRESUMEN
There is only scarce information available on genotypic diversity of the Mycobacterium tuberculosis complex (MTBC) clinical isolates circulating in the Northern part of Brazil, a relatively neglected region regarding research on tuberculosis. We therefore characterized 980 MTBC clinical isolates from the state of Pará, by spoligotyping and data was compared with patterns from around the world, besides analyzing drug susceptibility, and collecting sociodemographic data. We also performed 24 loci MIRU-VNTR typing to evaluate phylogenetic inferences among the East-African-Indian (EAI) lineage strains. The Geographic Information System analyses were performed to generate a descriptive visualization of MTBC strain distribution in the region. A total of 249 different spoligopatterns primarily belonging to evolutionary recent Euro-American lineages, as well as Central-Asian, Manu and ancestral EAI lineages, were identified, in addition to strains with reportedly unknown lineage signatures. The most frequent lineages were Latin American Mediterranean, T and Haarlem. Interestingly, EAI lineage strains were found in a significantly higher proportion in comparison with previous studies from South America. Regarding EAI lineage, the absence of spacers 4-9 and 23-24 co-related to 24 loci MIRU-VNTRs may suggest a close evolutionary relationship between such strains in Pará and those prevalent in Mozambique, which might have contributed to the genetic diversity of MTBC strains in this region.
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Variación Genética , Mycobacterium tuberculosis/clasificación , Mycobacterium tuberculosis/genética , Tuberculosis/epidemiología , Tuberculosis/microbiología , Adolescente , Adulto , Distribución por Edad , Brasil/epidemiología , Niño , Preescolar , ADN Bacteriano , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Repeticiones de Minisatélite , Tipificación Molecular , Filogenia , Filogeografía , Adulto JovenRESUMEN
BACKGROUND: Human papillomavirus (HPV) infection is associated with cervical cancer, the most frequent cancer in women from northern Brazil. Assessment of the short-term impact of HPV vaccination depends on the availability of data on the prevalence of type-specific HPV in young women in the pre-immunization period, although these data are currently unavailable for the study region. The aim of this study was to estimate the distribution of all mucosal HPV genotypes, including low- and high-risk HPV types, in unvaccinated college students from northern Brazil. FINDINGS: Specimens were collected from 265 university students during routine cervical cancer screening. The HPV DNA was assessed by Polymerase Chain Reaction and positive samples were genotyped by Restriction Fragment Length Polymorphism. Most students (85.7 %) had normal cytological results. The prevalence of HPV was 25.3 % (67/265), with a high frequency of multiple infections and non-vaccine high-risk HPV genotypes. The most prevalent type was HPV-61 (5.3 %), followed by types 82, 16, 59, and 6. Multiple infections were associated with high-risk and possibly high-risk HPVs. CONCLUSIONS: We demonstrated a high prevalence of HPV infection in university students from northern Brazil. Vaccine high-risk types were relatively rare, emphasizing the predominance of carcinogenic genotypes that are not prevented by the currently available vaccines. Our study highlights the need to reinforce cytological screening in women from northern Brazil, and promote the early diagnosis and treatment of the precancerous lesions associated with cervical cancer.
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BACKGROUND: HTLV-1 is a retrovirus that causes lymphoproliferative disorders and inflammatory and degenerative diseases of the central nervous system in humans. The prevalence of this infection is high in parts of Brazil and there is a general lack of public health care programs. As a consequence, official data on the transmission routes of this virus are scarce. OBJECTIVE: To demonstrate familial aggregation of HTLV infections in the metropolitan region of Belém, Pará, Brazil. METHOD: A cross-sectional study involving 85 HTLV carriers treated at an outpatient clinic and other family members. The subjects were tested by ELISA and molecular methods between February 2007 and December 2010. RESULTS: The prevalence of HTLV was 43.5% (37/85) for families and 25.6% (58/227) for the family members tested (95% CI: 1.33 to 3.79, Pâ=â0.0033). Sexual and vertical transmission was likely in 38.3% (23/60) and 20.4% (29/142) of pairs, respectively (95% CI: 1.25 to 4.69, Pâ=â0.0130). Positivity was 51.3% (20/39) and 14.3% (3/21) in wives and husbands, respectively (95% CI: 0.04 to 0.63, Pâ=â0.0057). By age group, seropositivity was 8.0% (7/88) in subjects <30 years of age and 36.7% (51/139) in those of over 30 years (95% CI: 0.06 to 0.34, P<0.0001). Positivity was 24.1% (7/29) in the children of patients infected with HTLV-2, as against only 5.8% (4/69) of those infected with HTLV-1 (95% CI: 0.05 to 0.72, Pâ=â0.0143). CONCLUSION: The results of this study indicate the existence of familial aggregations of HTLV characterized by a higher prevalence of infection among wives and subjects older than 30 years. Horizontal transmission between spouses was more frequent than vertical transmission. The higher rate of infection in children of HTLV-2 carriers suggests an increase in the prevalence of this virus type in the metropolitan region of Belém.
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Infecciones por Deltaretrovirus/epidemiología , Salud de la Familia , Virus Linfotrópico T Tipo 1 Humano/aislamiento & purificación , Virus Linfotrópico T Tipo 2 Humano/aislamiento & purificación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Brasil/epidemiología , Niño , Preescolar , Enfermedades Transmisibles Emergentes/epidemiología , Enfermedades Transmisibles Emergentes/transmisión , Estudios Transversales , Infecciones por Deltaretrovirus/transmisión , Transmisión de Enfermedad Infecciosa , Femenino , Humanos , Lactante , Transmisión Vertical de Enfermedad Infecciosa , Masculino , Persona de Mediana Edad , Enfermedades Desatendidas/epidemiología , Estudios Seroepidemiológicos , Adulto JovenRESUMEN
Noroviruses are the leading cause of epidemic, non-bacterial outbreaks of acute gastroenteritis, and are also a major cause of sporadic acute gastroenteritis in infants. The aim of the present study was to identify norovirus infections in children not infected by rotavirus admitted to hospital for acute gastroenteritis in Belém. A total of 348 fecal specimens were obtained from children with diarrhea aged less than 5 years, all of whom had tested negative for rotavirus, between May 2008 and April 2010. Fecal samples were screened for norovirus antigen using enzyme-immunoassay (EIA). Specimens were subjected to reverse-transcription polymerase chain reaction (RT-PCR) using the primers Mon432/434-Mon431/433 for detection of the GI and GII norovirus strains, respectively. Based on both methods, the overall norovirus positivity rate was 36.5% (127/348). Of the 169 samples collected in the first year, 44.4% (n = 75) tested positive for norovirus using both methods, 35.5% (n = 60) by EIA and 40.8% (n = 69) by RT-PCR. Using RT-PCR as a reference standard, a sensitivity of 78.3%, specificity of 94%, and agreement of 87.6% were recorded. Genome sequencing was obtained for 22 (31.9%) of the 69 positive samples, of which 90.9% (20/22) were genotype GII.4d and 9.1% (2/22) were genotype GII.b. Norovirus infection was most frequent in children under 2 years of age (41.5%-115/277). The peak incidence (62.1%) of norovirus-related acute gastroenteritis in these patients (not infected by rotavirus) was observed in February 2010. These findings emphasize the importance of norovirus as a cause of severe acute gastroenteritis among children in Belém, Pará, Northern Brazil.
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Infecciones por Caliciviridae/virología , Gastroenteritis/virología , Norovirus/clasificación , Norovirus/genética , Brasil/epidemiología , Infecciones por Caliciviridae/epidemiología , Preescolar , Ensayo de Inmunoadsorción Enzimática/métodos , Heces/virología , Femenino , Gastroenteritis/epidemiología , Genotipo , Hospitalización , Humanos , Lactante , Recién Nacido , Masculino , Norovirus/aislamiento & purificación , Prevalencia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Sensibilidad y EspecificidadRESUMEN
This study utilized the hsp65 polymerase chain reaction restriction analysis (PRA) method in the identification of nontuberculous mycobacteria (NTMs) isolated in a Brazilian mycobacteria laboratory. NTM isolates from clinical specimens collected from 192 patients were characterized using the hsp65 PRA method and analyzed using both 16S rRNA and hsp65 gene sequencing. Only 30% of the NTM strains were correctly identified through PRA, though the suggested inclusion of an additional restriction enzyme could increase the resolution to roughly 90%. A total of 17 NTM strains were not identified to species level and may represent a new taxonomic entity classified as belonging to the Mycobacterium simiae complex. This study demonstrates the applicability of hsp65 PRA in the identification of several NTM strains in a reference laboratory, though the results suggest that some modifications to the original PRA method could increase its resolution substantially.
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Infecciones por Mycobacterium/microbiología , Mycobacterium/clasificación , Mycobacterium/genética , Reacción en Cadena de la Polimerasa/métodos , Mapeo Restrictivo/métodos , Proteínas Bacterianas/genética , Técnicas de Tipificación Bacteriana/métodos , Brasil , Chaperonina 60/genética , ADN Bacteriano/análisis , Humanos , Mycobacterium/aislamiento & purificación , ARN Ribosómico 16S/genética , Estándares de Referencia , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
We isolated 44 strains of rapidly growing mycobacteria (RGM) from 19 patients with pulmonary infections assisted at the Instituto Evandro Chagas (Pará, Brazil) from 2004 to 2007. Identification at the species level was performed by PCR restriction fragment length polymorphism analysis (PRA) of a 441 bp hsp65 fragment and partial 16S rRNA, hsp65, and rpoB gene sequencing. Genotyping by PRA yielded 3 digestion patterns: one identical to Mycobacterium abscessus type I (group I); another to M. abscessus type II, Mycobacterium bolletii, and Mycobacterium massiliense (group II); and a third typical for Mycobacterium fortuitum type I (group III). When comparing analysis of the 3 genes, more discrimination was obtained by rpoB gene sequence, which allowed good distinction between group I, II, and III strains and subclassification of group II strains in SG IIa (M. bolletii) and SG IIb (M. massiliense). In this study, we show that the description of new RGM species requires the establishment of standardized procedures for RGM identification and the alert of the clinician about their involvement in pulmonary disease and the necessity of treatment for control and cure.