Short communication: HIV type 1 tropism determination in a novel dried blood spot membrane and the use of a mixture of outer nested polymerase chain reaction primers.

Genotropism was determined in 608 Brazilian samples collected in dried blood spots using Polyethersulfone collection cards. Patients were infected by subtype B (88.8%), F (5.6%), C (3.3%), A (1.8%), and G (0.5%). All patients were exposed to three classes of antiretrovirals, and 59.8% of the samples harbored R5 viruses, 35% non-R5-tropic viruses, and 5.1% harbored mixtures of R5 and non-R5-tropic viruses, with non-R5 more prevalent among clade B-infected patients as compared to non-B (42.8% versus 19.1%; p<0.0003). A strategy using a mixture of outer nested polymerase chain reaction (PCR) primers reduced the number of negative PCR results from 39% to 19%.

The use of real-time PCR to detect hepatitis C virus RNA in dried blood spots from Brazilian patients infected chronically.

Collecting and transporting samples for RNA analysis can be challenging, especially in situations where financial resources are limited. In this study, a quantitative real-time PCR (qPCR) for the analysis of HCV RNA was developed and adapted for use with dried blood spot (DBS) samples. A qPCR for HCV 5'NCR, an internal control and a calibration curve were developed, and the sensitivity, specificity and dynamic range of amplification were evaluated using a panel of viruses. Plasma and DBS samples from 100 patients who had completed four weeks of Peginterferon alfa-2b+Ribavirin treatment were collected (DBS on SS903 collection cards and transported at room temperature). After 24 weeks of treatment, samples were collected from 68 of these patients. Of the 168 samples, 2 yielded false-negative results, and 4 yielded false-positive results (sensitivity was 98%, specificity was 94.3%, positive predictive value was 96.1%, and negative predictive value was 96.9%). Additionally, 2039 DBS samples from 1114 patients currently undergoing treatment for a chronic HCV infection in a clinical trial were tested. Only 10 samples out of the 2039 yielded invalid results warranting re-collection of DBS. The detection of HCV RNA in DBS can be a cost-effective strategy for HCV treatment monitoring, especially in settings where resources are limited.

Comparison of direct immunofluorescence, conventional cell culture and polymerase chain reaction techniques for detecting respiratory syncytial virus in nasopharyngeal aspirates from infants.

A total of 316 samples of nasopharyngeal aspirate from infants up to two years of age with acute respiratory-tract illnesses were processed for detection of respiratory syncytial virus (RSV) using three different techniques: viral isolation, direct immunofluorescence, and PCR. Of the samples, 36 (11.4%) were positive for RSV, considering the three techniques. PCR was the most sensitive technique, providing positive findings in 35/316 (11.1%) of the samples, followed by direct immunofluorescence (25/316, 7.9%) and viral isolation (20/315, 6.3%) (p < 0.001). A sample was positive by immunofluorescence and negative by PCR, and 11 (31.4%) were positive only by RT-PCR. We conclude that RT-PCR is more sensitive than IF and viral isolation to detect RSV in nasopharyngeal aspirate specimens in newborn and infants.

Comparison of direct immunofluorescence, conventional cell culture and polymerase chain reaction techniques for detecting respiratory syncytial virus in nasopharyngeal aspirates from infants / Comparação entre imunofluorescência direta, cultura convencional de células e reação em cadeia de polimerase para detectar vírus respiratório sincicial em lavados de nasofaringe de crianças

A total of 316 samples of nasopharyngeal aspirate from infants up to two years of age with acute respiratory-tract illnesses were processed for detection of respiratory syncytial virus (RSV) using three different techniques: viral isolation, direct immunofluorescence, and PCR. Of the samples, 36 (11.4 percent) were positive for RSV, considering the three techniques. PCR was the most sensitive technique, providing positive findings in 35/316 (11.1 percent) of the samples, followed by direct immunofluorescence (25/316, 7.9 percent) and viral isolation (20/315, 6.3 percent) (p < 0.001). A sample was positive by immunofluorescence and negative by PCR, and 11 (31.4 percent) were positive only by RT-PCR. We conclude that RT-PCR is more sensitive than IF and viral isolation to detect RSV in nasopharyngeal aspirate specimens in newborn and infants.

Um total de 316 amostras de lavado de nasofaringe obtidas de crianças em acompanhamento ambulatorial com até dois anos de idade durante episódio de doença aguda do trato respiratório foram processadas para detecção do vírus sincicial respiratório (VSR) utilizando três diferentes técnicas: isolamento viral, imunofluorescência direta e reação em cadeia por polimerase (RT-PCR). Destas amostras, 36 (11,4 por cento) foram positivas para o VSR. A RT-PCR foi a técnica mais sensível, com positividade em 35 (11,1 por cento) das amostras, seguindo-se a imunofluorescência direta (25/316, 7,9 por cento) e o isolamento viral (20/315, 6,3 por cento) (p < 0,001). Uma amostra foi positiva pela imunofluorescência e negativa pela RT-PCR, e 11/36 (31,4 por cento) foram positivas somente pela RT-PCR. Concluímos que a RT-PCR é mais sensível que a imunofluorescência e o isolamento viral para detecção do VRS em amostras de aspirado de nasofaringe de recém-nascidos e lactentes.

Seroprevalence of antibodies against varicella-zoster virus and response to the varicella vaccine in pediatric renal transplant patients.

The severity of varicella-zoster virus (VZV) in immunocompromised children, especially in those receiving renal transplants, is well known. However, the use of live attenuated virus vaccine in this population is controversial. This study aimed to: (i) assess the immunization status of pediatric renal transplant recipients at our center; (ii) determine the anti-VZV antibody titers in such patients; (iii) evaluate the response to VZV vaccine in seronegative children and in those who present low antibody titers (defined as <500 mAU/mL). Vaccinated children were monitored for adverse effects for 8 wk after vaccination. Fifty patients with a mean age of 13.7 yr (range, 3-17 yr) were enrolled. In 49, blood samples were collected and antibodies were screened using ELISA. Seropositivity to VZV was found in 43 (88%), and antibody titers were >/=500 mAU/mL in 37 (75.5%). Of the 12 children who were eligible for vaccination and had antibody titers <500 mAU/mL, one developed varicella before vaccination, two did not meet the inclusion criteria, and three parents refused the vaccination. In the six vaccinated children, there were no adverse reactions to the vaccine, and four (66.6%) responded with anti-VZV titers >/=500 mAU/mL 6-8 wk after vaccination. In conclusion, after renal transplantation, varicella vaccine is safe with a 66% rate of conversion to high antibody titers.

[Prevalence of varicella-zoster virus antibodies in young adults from different Brazilian climatic regions]. / Prevalência de anticorpos para o vírus da varicela-zoster em adultos jovens de diferentes regiões climáticas brasileiras.

To evaluate the prevalence of varicella-zoster virus infection in young adults from different Brazilian urban regions, 975 serum samples from blood donors aged 20 to 29 years, from tropical climate cities (Salvador and Fortaleza) and from temperate climate cities (S o Paulo, Curitiba and Porto Alegre) were tested by an in-house ELISA for detection of anti-varicella-zoster virus IgG antibodies. The overall prevalence was 94.2%. The lowest rate was observed in Fortaleza (88.7%) and the highest in Curitiba (99.5%). Seroprevalence in tropical regions of Brazil (89.4%) was significantly higher than in temperate regions (97.3%), a similar pattern to that observed in other tropical countries.

Prevalência de anticorpos para o vírus da varicela-zoster em adultos jovens de diferentes regiöes climáticas brasileiras / Prevalence of varicella-zoster virus antibodies in young adults from different Brazilian climatic regions

Para avaliar a prevalência da infecçäo pelo vírus da varicela-zoster, de regiöes urbanas de diferentes regiöes do Brasil, 975 amostras de soro provenientes de adultos jovens doadores de sangue com idade entre 20 e 29 anos, de cidades de clima tropical (Salvador e Fortaleza) e de clima temperado (São Paulo, Curitiba e Porto Alegre) foram processadas pelo teste imunoenzimático doméstico para pesquisa de anticorpos IgG anti-Vírus da varicela zoster. A soroprevalência global de anticorpos anti-virus da varicela zoster nas várias regiöes estudadas foi de 94,2 por cento. A menor taxa (88,7 por cento) foi observada em Fortaleza e a maior em Curitiba (99,5 por cento). A soroprevalência nas regiöes de clima tropical (89,4 por cento) foi significativamente inferior a soroprevalência nas regiöes de clima temperado (97,3 por cento), seguindo um padräo similar à infecçäo em outros países de clima tropical