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.

Antiretroviral drug resistance among patients with human immunodeficiency virus who act as sources or potential sources in occupational accidents involving healthcare workers.

OBJECTIVE: To determine human immunodeficiency virus (HIV) type 1 genotypic antiretroviral drug resistance profiles of patients presenting a risk or potential risk for occupational exposure of healthcare workers. DESIGN: Observational survey involving HIV-infected patients. Blood samples collected from source-patients and potential source-patients underwent HIV-1 genotypic antiretroviral resistance testing and determination of CD4 counts and viral load. Affected healthcare workers were monitored for 6 months after exposure. SETTING: The survey was conducted in a tertiary-care hospital located in Sao Paulo, Brazil. Sao Paulo is considered the epicenter of the HIV-acquired immunodeficiency (AIDS) virus epidemic in Brazil. PARTICIPANTS: Source-patients, potential source-patients, and affected healthcare workers. RESULTS: A total of 371 occupational exposures to biological materials were reported, 46 (12.3%) of which were from HIV-seropositive source-patients. Samples from 18 source-patients and 26 patients considered "potential sources for accidents" were analyzed. Of these 44 samples, 18 (41%) presented resistance-related mutations in reverse transcriptase, protease, or both. Of these 18 samples, 16 (89%) had resistance to drugs included in the prophylactic schedule recommended by the Brazilian Ministry of Health. CONCLUSIONS: Use of the Centers for Disease Control and Prevention-Brazilian post-exposure prophylaxis regimen will result in the administration of antiretroviral agents to which the source HIV-1 isolate will often be resistant. Therefore, it would be advisable to carefully investigate the history of use of antiretroviral agents by source-patients and adjust the prophylactic therapy based on those data and, subsequently, the results of resistance testing.

Cut and puncture accidents involving health care workers exposed to biological materials

The first repor of occupational acquisition of HIV appeared in 1984, and, by june,1997, the Centers for Disease Control and Prevention (CDC) had reported 52 document cases of sero-conversion following occupational exposure to HIV-1 by health care workers of those cases. 47(90.3 percent) were exposed to blood. The most frequent type of accident reported was percutaneous needlestick injury. Prospective studies have estimated that the risk of HIV transmission following percutaneous exposure to infected blood is 0,3 percent (Confidence Interval 95 percent=0,2 percent to 0,5 percent). Following a mucous membrane exposure, the risk is 0,09 percent (CI95 percent=0.006 percent to 0,5 percent). The risk of hepatitis B acquisition ranges from 6 percent to 30 percent, and hepatitis C acquisition, 3 percent to 10 percent. Since 1992, the São Paulo Hospital's Hospital Infectious Prevention and Control Service (SPCIH) has notified and treated all workers exposed to accidents involving biological materials. In the last six years, we have handled approximately 1,300 cases of reported accidents, of which 90 percent were percutaneous, most involving needlesticks. Such cases were frequently caused by the inadequate disposal and reccaping of needles. In these accidents, 20 percent of the source patients were HIV positive, 10 percent were hepatitis C positive, and 7,6 percent were hepatitis B positive. This review summarizes the guidelines for a standardized response When deadling with accidents involving health care workers. Transmission of hepatitis B and HIV can be reduced if adequate preventive measures are taken in advance. If proper prophyllaxis is not being done, it should be initiated immediately.

Meningoencefalite na fase aguda do sarampo: relato de seis casos / Meningoencephalitis in the acute phase of measles: report of 6 cases

Apresentamos 6 casos de meningoencefalite aguda por sarampo, diagnosticados durante epidemia na cidade de São Paulo, em 1997. Os prontuários dos 6 pacientes foram analisados retrospectivamente. O diagnóstico de meningoencefalite baseou-se nas alterações clínicas e liquóricas, e foi confirmado por sorologia específica. Dos 467 pacientes com sarampo atendidos no Instituto de Infectologia Emílio Ribas nesse período, estes 6 evoluíram com alterações neurológicas e liquóricas durante a fase exantemática, sendo a sonolência e rigidez de nuca os achados mais frequentes. Os pacientes tinham entre 2 meses e 28 anos de idade. O exame do líquor mostrou pleocitose em todos. Não houve correlação entre a severidade do quadro clínico e liquórico com a evolução. Em 4 casos foi necessária internação na unidade de terapia intensiva; destes, 2 foram intubados. Apenas 2 pacientes apresentaram alterações na tomografia computadorizada de crânio. Todos tiveram boa evolução, sem sequelas.