Diagnostic accuracy and applicability of a PCR system for the detection of Schistosoma mansoni DNA in human urine samples from an endemic area.
Schistosomiasis caused by Schistosoma mansoni, one of the most neglected human parasitoses in Latin America and Africa, is routinely confirmed by microscopic visualization of eggs in stool. The main limitation of this diagnostic approach is its lack of sensitivity in detecting individual low worm burdens and consequently data on infection rates in low transmission settings are little reliable. According to the scientific literature, PCR assays are characterized by high sensitivity and specificity in detecting parasite DNA in biological samples. A simple and cost effective extraction method for DNA of Schistosoma mansoni from urine samples in combination with a conventional PCR assay was developed and applied in an endemic area. This urine based PCR system was tested for diagnostic accuracy among a population of a small village in an endemic area, comparing it to a reference test composed of three different parasitological techniques. The diagnostic parameters revealed a sensitivity of 100%, a specificity of 91.20%, positive and negative predictive values of 86.25% and 100%, respectively, and a test accuracy of 94.33%. Further statistical analysis showed a k index of 0.8806, indicating an excellent agreement between the reference test and the PCR system. Data obtained from the mouse model indicate the infection can be detected one week after cercariae penetration, opening a new perspective for early detection and patient management during this stage of the disease. The data indicate that this innovative PCR system provides a simple to handle and robust diagnostic tool for the detection of S. mansoni DNA from urine samples and a promising approach to overcome the diagnostic obstacles in low transmission settings. Furthermore the principals of this molecular technique, based on the examination of human urine samples may be useful for the diagnosis of other neglected tropical diseases that can be detected by trans-renal DNA.
Laboratório de Esquistossomose, Centro de Pesquisas René Rachou, FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil. marenk@cpqrr.fiocruz.br
Martin Johannes
MJ
Enk
Guilherme
G
Oliveira e Silva
Nilton Barnabé
NB
Rodrigues
D000818:Animals; D001938:Brazil / Q000453:epidemiology; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000652:urine*; D019353:Endemic Diseases; D005243:Feces / Q000469:parasitology; D006801:Humans; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods; D011237:Predictive Value of Tests; D012550:Schistosoma mansoni / Q000235:genetics*; D012555:Schistosomiasis mansoni / Q000175:diagnosis* / Q000453:epidemiology*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018509:DNA, Helminth