Prevalence of Treponema pallidum DNA among blood donors with two different serologic tests profiles for syphilis in São Paulo, Brazil.

The presence of Treponema pallidum DNA was assessed by real-time PCR in samples of blood donors with reactive serologic tests for syphilis. Treponema pallidum DNA was detected in two (1·02%) of 197 samples of VDRL>8, EIA+ and FTA-ABS+ donors, and in no sample from 80 VDRL−, EIA+ and FTA-ABS+ donors. Donors VDRL−, EIA+ and FTA-ABS+ lack demonstrable T. pallidum DNA in their blood and are unlike to transmit syphilis. Donors VDRL>8, EIA+ and FTA-ABS+ carry the risk of syphilis infectivity even in concomitance to antibodies detection. Serologic screening for syphilis may still play a role to prevent its transfusion transmission.

Demographic, risk factors and motivations among blood donors with reactive serologic tests for syphilis in São Paulo, Brazil.

OBJECTIVE: To identify the demographic characteristics, risk factors and motivations for donating among blood donors with reactive serologic tests for syphilis. BACKGROUND: Post-donation interviews with syphilis seropositive blood donors improve recruitment and screening strategies. METHODS: This case-control study compares 75 Venereal Disease Research Laboratory (VDRL) > 8, EIA+ (enzyme immunoassay) and FTA-ABS+ (fluorescent treponemal antibody); 80 VDRL-, EIA+ and FTA-ABS+; and 34 VDRL- and EIA- donors between 2004 and 2009. Donors were assessed by their demographic characteristics, sexual behaviour, history of alcohol and illicit drugs use, and motivations to donate. RESULTS: Donors with VDRL > 8 were more likely to be divorced [AOR = 12·53; 95% confidence interval (CI) 1·30-120·81], to have had more than six sexual partners (AOR=7·1; 95% CI 1·12-44·62) and to report male-male-sex in the past 12 months (AOR=8·18; 95% CI 1·78-37·60). Donors with VDRL-, EIA+ and FTA-ABS+ were less likely to be female (AOR=0·26; 95% CI 0·07-0·96), more likely to be older (AOR=10·2; 95% CI 2·45-42·58 ≥ 39 and <60 years old) and to have had more than six sexual partners in the past 12 months (AOR = 8·37; 95% CI 1·49-46·91). There was no significant difference among groups regarding illicit drugs use; 30·7% (VDRL > 8) and 12·5% (VDRL-, EIA+ and FTA-ABS+) of donors reported that they had been at risk for HIV infection (P = 0·004). One-third of donors came to the blood bank to help a friend or a relative who needed blood. CONCLUSION: Although donors exposed to syphilis reported and recognised some high risk behaviour, most were motivated by direct appeal to donate blood. Monitoring the risk profile of blood donors can benefit public health and improve blood safety.

Cost-effective analysis of different algorithms for the diagnosis of hepatitis C virus infection.

We compared the cost-benefit of two algorithms, recently proposed by the Centers for Disease Control and Prevention, USA, with the conventional one, the most appropriate for the diagnosis of hepatitis C virus (HCV) infection in the Brazilian population. Serum samples were obtained from 517 ELISA-positive or -inconclusive blood donors who had returned to Fundação Pró-Sangue/Hemocentro de São Paulo to confirm previous results. Algorithm A was based on signal-to-cut-off (s/co) ratio of ELISA anti-HCV samples that show s/co ratio > or =95% concordance with immunoblot (IB) positivity. For algorithm B, reflex nucleic acid amplification testing by PCR was required for ELISA-positive or -inconclusive samples and IB for PCR-negative samples. For algorithm C, all positive or inconclusive ELISA samples were submitted to IB. We observed a similar rate of positive results with the three algorithms: 287, 287, and 285 for A, B, and C, respectively, and 283 were concordant with one another. Indeterminate results from algorithms A and C were elucidated by PCR (expanded algorithm) which detected two more positive samples. The estimated cost of algorithms A and B was US$21,299.39 and US$32,397.40, respectively, which were 43.5 and 14.0% more economic than C (US$37,673.79). The cost can vary according to the technique used. We conclude that both algorithms A and B are suitable for diagnosing HCV infection in the Brazilian population. Furthermore, algorithm A is the more practical and economical one since it requires supplemental tests for only 54% of the samples. Algorithm B provides early information about the presence of viremia.

Cost-effective analysis of different algorithms for the diagnosis of hepatitis C virus infection

We compared the cost-benefit of two algorithms, recently proposed by the Centers for Disease Control and Prevention, USA, with the conventional one, the most appropriate for the diagnosis of hepatitis C virus (HCV) infection in the Brazilian population. Serum samples were obtained from 517 ELISA-positive or -inconclusive blood donors who had returned to Fundação Pró-Sangue/Hemocentro de São Paulo to confirm previous results. Algorithm A was based on signal-to-cut-off (s/co) ratio of ELISA anti-HCV samples that show s/co ratio ≥95 percent concordance with immunoblot (IB) positivity. For algorithm B, reflex nucleic acid amplification testing by PCR was required for ELISA-positive or -inconclusive samples and IB for PCR-negative samples. For algorithm C, all positive or inconclusive ELISA samples were submitted to IB. We observed a similar rate of positive results with the three algorithms: 287, 287, and 285 for A, B, and C, respectively, and 283 were concordant with one another. Indeterminate results from algorithms A and C were elucidated by PCR (expanded algorithm) which detected two more positive samples. The estimated cost of algorithms A and B was US$21,299.39 and US$32,397.40, respectively, which were 43.5 and 14.0 percent more economic than C (US$37,673.79). The cost can vary according to the technique used. We conclude that both algorithms A and B are suitable for diagnosing HCV infection in the Brazilian population. Furthermore, algorithm A is the more practical and economical one since it requires supplemental tests for only 54 percent of the samples. Algorithm B provides early information about the presence of viremia.

Prevalence of GB virus C (hepatitis G virus) and risk factors for infection in São Paulo, Brazil.

The prevalence of GB virus C (GBV-C) varies widely throughout the world. A cross-sectional study was conducted in the city of São Paulo, Brazil, to estimate the prevalence of GBV-C infection and to identify associated risk factors, using a large sampling of the general population rather than blood donors or an illness-related group of subjects. GBV-C RNA was detected by reverse-transcriptase polymerase chain reaction using primers directed to the 5' noncoding region (NCR) and nonstructural 5A region (NS5A) in serum samples from 1,039 healthy individuals 2 years of age or more. Fifty-two individuals were positive for both sets of primers and one was positive for NS5A only (prevalence of GBV-C infection, 5.1%; 95%CI, 3.9-6.7%). No child under 5 years of age was found positive. Among subjects aged 5 years or more, the prevalence of infection increased consistently with age, up to 30-39 years (8.3%), and decreased from then on. The number of sexual partners in the last 3 years (2 or more: OR, 2.6; 95%CI, 1.3-5.5) and history of contact with blood-sucking insects (OR, 2.5; 95%CI 1.2-5.4) were independently associated with GBV-C infection. In conclusion, the prevalence of GBV-C infection is high in São Paulo. In addition to parenteral transmission, another route, e.g. sexual or vertical, may be involved.

An improved, PCR-based strategy for the detection of Trypanosoma cruzi in human blood samples.

Attempts were made to improve the PCR-based detection of Trypanosoma cruzi in blood samples, primarily for screening blood donors. Samples were obtained from candidate donors who were reactive in one or two of three serological tests for Chagas disease (and therefore considered 'indeterminate') or in all three tests (3+). Each sample was then examined using three different, PCR-based techniques: 'PCR-I' (in which the target DNA is a nuclear repetitive sequence); 'PCR-II' [amplifying a conserved region of the T. cruzi kinetoplast DNA (kDNA)]; and 'PCR-III' (a new strategy in which the target kDNA is amplified by 'nested' PCR). Among the samples from 3+ individuals, PCR-I, PCR-II and PCR-III amplified two (3.8%) out of 52, four (4.5%) out of 88, and 27 (25.7%) out of 105 samples tested, respectively. Seven, 69 and 70 samples from 'indeterminate' subjects were tested by PCR-I, PCR-II and PCR-III, respectively; there was not a single positive result by PCR-I or PCR-II, but three (4.3%) of the samples tested by PCR-III were positive. In a reconstruction experiment, in conditions in which PCR-I and PCR-II could not detect 10,000 parasites/ml, PCR-III was able to detect one parasite/ml. Although all three PCR-based strategies examined had rather poor sensitivities, PCR-III was far more sensitive than PCR-I or PCR-II.