First detection of hepatitis E virus in Central Argentina: environmental and serological survey.

BACKGROUND: The hepatitis E virus (HEV) is an emergent causative agent of acute hepatitis worldwide, transmitted by fecal-oral route. In Argentina it is considered rare, so differential laboratory testing is not routinely performed. Besides, in Argentina's central area epidemiological and molecular characteristics of HEV are still unknown. OBJECTIVES: Provide evidence of local circulation of HEV by molecular detection on environmental samples and by serological survey in healthy adult population of Córdoba city, Argentina. STUDY DESIGN: Environmental surveillance was conducted in river and sewage samples collected between 2007 and 2009-2011. Viral detection was performed by RT-Nested PCR of ORF-1 and ORF-2 partial regions. Anti-HEV IgG was determined by EIA in 433 serum samples collected between 2009 and 2010. RESULTS: HEV was detected in 6.3% of raw sewage samples and in 3.2% of riverine samples. Nucleotide sequencing analyses revealed that all isolates belonged to genotype 3, subtypes a, b and c. The prevalence of IgG anti-HEV was 4.4%. Seroprevalence increased with the age of the individuals (OR: 3.50; 95% CI 1.39-8.87; p=0.0065) and, although the prevalence was higher in low income population, no statistical relation was found between anti-HEV and socioeconomic level. CONCLUSIONS: The environmental findings added to serological results, demonstrate that HEV circulates in central Argentina. Contamination of water with HEV could represent a route of transmission for local populations, which have a high number of susceptible individuals. This fact alerts local health care systems in order to include detection of HEV in the diagnostic algorithm of viral hepatitis.

Distribution of polymorphisms in cytochrome P450 2B6, histocompatibility complex P5, chemokine coreceptor 5, and interleukin 28B genes in inhabitants from the central area of Argentina.

AIMS: The selection of the most appropriate treatment for several diseases relies on a number of factors such as environment, age, gender, and nutrition. Additionally, the contribution of different genetic polymorphisms to treatment efficacy has been largely recognized. The lack of information on the pharmacogenetic profile of our population prompted us to analyze the frequency of polymorphisms known to be relevant to achieve treatment efficacy with different therapeutic agents in viral infectious diseases, such as Hepatitis C and AIDS. RESULTS: The allelic frequencies for the wild-type variant of the genes analyzed were cytochrome P450 2B6 (CYP2B6; rs3745274; 516G) 0.618 (95% confidence interval [CI]: 0.523, 0.711), chemokine coreceptor 5 (CCR5; rs333) 0.961 (95% CI: 0.942, 0.98), histocompatibility complex P5 (HCP5; rs2395029; 335T) 0.971 (95% CI: 0.937, 1), and interleukin 28B (IL28B; rs12979860; 12007005C) 0.656 (95% CI: 0.564, 0.747), respectively. CONCLUSIONS: Our data indicate that the genetic profile of the population studied is similar to that reported for other Caucasian populations, with only slight differences for CYP2B6. Noteworthy, the considerable number of patients carrying CYP2B6 (516T) and IL28B (12007005T) alleles underlies the importance of considering pharmacogenetic testing before starting drug therapy protocols to prevent toxicity and/or lack of effectiveness in AIDS or hepatitis C virus infections.

Genetic profiling of GSTP1, DPYD, FCGR2A, FCGR3A and CCND1 genes in an Argentinian population.

OBJECTIVES: To determine the frequencies of relevant allelic variants in oncology for the GSTP1, DPYD, FCGR2A, FCGR3A and CCND1 genes in a population from Central Argentina. To compare the allelic distribution found with the frequencies reported for other ethnic groups. DESIGN AND METHODS: Genotyping was carried out in a total of 102 unrelated Argentinian subjects. FCGR3A (rs396991) was detected using allele specific polymerase chain reaction (PCR) assay, while GSTP1 (rs1695), DPYD (rs3918290), FCGR2A (rs1801274) and CCND1 (rs9344) variants were assessed by PCR-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The allele frequencies for GSTP*1B, DPYD*2A, FCGR2A (131R), FCGR3A (158F) and CCND1 (870G) in Argentinians were 0.35, 0.005, 0.41, 0.77 and 0.47, respectively. CONCLUSIONS: We found that the Argentinian population tested resembles other Caucasians populations, especially Spaniards; yet the differences in allele distribution with other Caucasian groups, uncover population admixture with native Amerindian and other ethnic groups, consistent with the well documented immigration flows landing Argentina from several countries.

Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples.

AIMS: Molecular biology techniques based on the detection of genomic sequences by reverse transcription combined with polymerase chain reaction (PCR) have enabled the detection of different RNA viruses in serum or plasma samples. Since the dengue epidemic outbreak declared in Argentina in 2009, numerous patients' samples were analyzed for the acute phase of infection. One of the main methodological drawbacks is the lack of internal control to measure the effectiveness of the viral extraction and reverse transcription process. In this article, we propose to standardize a molecular method to detect beta actin (ß-Act) and glucose 6 phosphate dehydrogenase (G6PDH) complementary DNAs (cDNAs) present in patient's plasma/serum, as a control process. RESULTS: RNA extraction, reverse transcription, and PCRs for human G6PDH, ß-Act, and the dengue virus genome were performed. cDNA fragments for ß-Act and G6PDH were amplified for all samples, regardless of the presence or absence of viral RNA. CONCLUSIONS: Amplification of ß-Act and G6PDH cDNAs can be used as a control for the extraction and reverse transcription processes during dengue virus detection. This could also be a useful method for controlling the above steps when infections caused by other RNA viruses are studied, even if another methodology is employed, such as real-time PCR.

Rapid HCP5 single-nucleotide polymorphism genotyping: a simple allele-specific PCR method for prediction of hypersensitivity reaction to Abacavir.

BACKGROUND: The most important factor limiting the success of an antiretroviral therapy is toxicity. The HLA-B*5701 allele is predictive of hypersensitivity reaction to Abacavir, and this gene is in a perfect linkage disequilibrium with the rs2395029 SNP present in the HCP5 gene. METHODS: Genomic DNA was extracted from blood obtained from 201 unrelated healthy Argentinean volunteers. The DNA was subjected to an allele-specific PCR method. Sequencing was performed to validate the test results. RESULTS: We were successful to amplify specific fragment of interest from the DNA samples. The method is easy, specific and reproducible. CONCLUSIONS: The application of this methodology is a rapid and simple method to detect the HCP5 polymorphism (rs2395029) previous to administration of Abacavir in patients with HIV infection.

Comparison of two serologic methods for the diagnosis of hydatidosis / Comparación de dos métodos serológicos para el diagnóstico de hidatidosis

El suero de 176 pacientes con antecedentes epidemiológicos o signos radiológicos y clínicos de hidatidosis fue sometido a contrainmunoelectroforesis (CIEF) e inmunoensayo enzimático (ELISA), utilizándose para ambas técnicas un antígeno semipurificado obtenido de los quistes de pacientes. Los resultados se compararon con los obtenidos por estudios radiológicos complementarios y se corroboraron mediante el examen de los quistes resecados. La biopsia confirmó el diagnóstico de hidatidosis en 65 (37%) pacientes y reveló la presencia de otras enfermedades en los 111 (63%) pacientes restantes. De los 176 pacientes estudiados, 36 (20,4%) tuvieron resultados positivos en la CIEF y 62 (35,2%) en el ELISA. Los resultados de ambas técnicas mostraron una excelente correlación con el diagnóstico posquirúrgico, ya que no hubo un solo resultado positivo falso y el ELISA produjo resultados negativos falsos en tres (4,6%) pacientes con quistes infectados, infértiles o con algún grado de calcificación. Por último se describe la estandarización de un microELISA fácil de usar y barato.

The sera of 176 patients with epidemiologic antecedents or radiologic and clinical signs of hydatidosis were tested by counterimmunoelectrophoresis (CIE) and enzyme-linked immunoassay (ELISA). A semipurified antigen from cysts of human origin was used for both techniques. The results were compared with those obtained from complementary radiologic studies and were confirmed by examination of excised cysts. Biopsy confirmed the diagnosis of hydatidosis in 65 patients (37%) and revealed the presence of other diseases in the remaining 111 (63%). Of the original 176 patients, 36 (20.4%) were positive by CIE and 62 (35.2%) by ELISA. Both techniques showed an excellent correlation with postsurgical diagnosis; neither produced any false positives, and the ELISA gave false negative results for only three patients (4.6%) with cysts that were infected, infertile, or calcified to some degree. The paper describes standardization of an inexpensive and easy-to-use microELISA.