Evaluation of real-time PCR assay to detect Schistosoma mansoni infections in a low endemic setting

Background\ud Schistosomiasis constitutes a major public health problem, and 200 million people are estimated to be infected with schistosomiasis worldwide. In Brazil, schistosomiasis has been reported in 19 states, showing areas of high and medium endemicity and a wide range of areas of low endemicity (ALE). Barra Mansa in Rio de Janeiro state has an estimated prevalence of 1%. ALE represent a new challenge for the helminth control because about 75% of infected individuals are asymptomatic and infections occur with a low parasite load (<100 eggs per gram of feces), causing a decrease in sensitivity of stool parasitological techniques, which are a reference for the laboratory diagnosis of this helminth. The objective of this study was to evaluate the performance of a TaqMan quantitative polymerase chain reaction (qPCR) technique in serum and feces DNA samples using the techniques of Kato-Katz (KK), Hoffman, Pons and Janer (HH) as references, during an epidemiological survey using fecal samples and sera from randomized residents from an ALE.\ud \ud \ud Methods\ud A cross-sectional study conducted from April to December 2011 using a probabilistic sampling that collected 572 fecal and serum samples. The laboratory diagnostic techniques used were: KK, HH and qPCR (feces and serum).\ud \ud \ud Results\ud We obtained the following results using the different diagnostic techniques: KK and HH, 0.9% (n =5); qPCR-feces, 9.6% (n =55); and qPCR-serum, 1.4% (n =8). The qPCR-feces presented the highest positivity, whereas the techniques of HH and KK were the least sensitive to detect infections (0.8%). Compared to HH and KK, qPCR-feces showed a statistically significant difference in positivity (p <0.05), although with poor agreement.\ud \ud \ud Conclusion\ud The positivity rate presented by the qPCR approach was far higher than that obtained by parasitological techniques. The lack of adequate surveillance in ALE of schistosomiasis indicates a high possibility of these areas being actually of medium and high endemicity. This study presents a control perspective, pointing to the possibility of using combined laboratory tools in the diagnosis of schistosomiasis in ALE.This study was supported by the São Paulo Research Foundation\ud (grants: 07/53457-7, 08/50461-6, and 10/52615-0). We would like to thank\ud Dr. Luciana Inácia Gomes and Dr. Ana Rabello, from the Laboratory of\ud Clinical Research of the René Rachou Research Institute - Fiocruz, Belo\ud Horizonte, Brazil, for their assistance during the standardization process.\ud We would also like to thank Ms. Magali Orban, from the Parasitology Section\ud of the Central Laboratory Division, Clinics Hospital, School of Medicine of the\ud University of São Paulo, for her technical assistance in conducting the\ud parasitological analysis of fecal samples and Ms. Maria Cristina Conceição de\ud Mello (LIM-06-FMUSP) for her excellent technical assistance concerning the\ud maintenance of the S. mansoni biological cycle. EDN and JRRP are research\ud Table 6 Agreement among the results obtained by the techniques KK and HH and those obtained by qPCR-serum and\ud qPCR-feces, in fecal and serum samples collected from the sampled population in the municipality of Barra\ud Mansa/RJ - 2011\ud Test KK-HH Total p McNemar Kappa CI (95%)\ud Negative Positive\ud N % n % n % Minimum Maximum\ud qPCR-feces\ud Negative 549 (90) 1 (0.2) 550 90.2