Is PCR the Next Reference Standard for the Diagnosis of Schistosoma in Stool? A Comparison with Microscopy in Senegal and Kenya

PLoS Negl Trop Dis. 2015 Jul 28;9(7):e0003959. doi: 10.1371/journal.pntd.0003959. eCollection 2015.

Abstract

Background: The current reference test for the detection of S. mansoni in endemic areas is stool microscopy based on one or more Kato-Katz stool smears. However, stool microscopy has several shortcomings that greatly affect the efficacy of current schistosomiasis control programs. A highly specific multiplex real-time polymerase chain reaction (PCR) targeting the Schistosoma internal transcriber-spacer-2 sequence (ITS2) was developed by our group a few years ago, but so far this PCR has been applied mostly on urine samples. Here, we performed more in-depth evaluation of the ITS2 PCR as an alternative method to standard microscopy for the detection and quantification of Schistosoma spp. in stool samples.

Methodology/principal findings: Microscopy and PCR were performed in a Senegalese community (n = 197) in an area with high S. mansoni transmission and co-occurrence of S. haematobium, and in Kenyan schoolchildren (n = 760) from an area with comparatively low S. mansoni transmission. Despite the differences in Schistosoma endemicity the PCR performed very similarly in both areas; 13-15% more infections were detected by PCR when comparing to microscopy of a single stool sample. Even when 2-3 stool samples were used for microscopy, PCR on one stool sample detected more infections, especially in people with light-intensity infections and in children from low-risk schools. The low prevalence of soil-transmitted helminthiasis in both populations was confirmed by an additional multiplex PCR.

Conclusions/significance: The ITS2-based PCR was more sensitive than standard microscopy in detecting Schistosoma spp. This would be particularly useful for S. mansoni detection in low transmission areas, and post-control settings, and as such improve schistosomiasis control programs, epidemiological research, and quality control of microscopy. Moreover, it can be complemented with other (multiplex real-time) PCRs to detect a wider range of helminths and thus enhance effectiveness of current integrated control and elimination strategies for neglected tropical diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Feces / parasitology*
  • Humans
  • Kenya / epidemiology
  • Microscopy / methods*
  • Polymerase Chain Reaction / methods*
  • Reference Standards
  • Schistosoma mansoni / isolation & purification*
  • Schistosomiasis mansoni / diagnosis*
  • Schistosomiasis mansoni / epidemiology
  • Senegal / epidemiology

Grants and funding

The Senegalese study received funding from the European Union’s sixth framework program (INCO-CT-2006-032405 SCHISTOINIR to KP and SM; http://cordis.europa.eu/fp6/). The Kenyan study received financial support from The University of Georgia Research Foundation through a grant from the Bill and Melinda Gates Foundation and also from USAID-supported NTD control activities at the CDC, Atlanta. The PCR analysis at Leiden University Medical Center was partly supported by the Prof. Dr. P.C. Flu-Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.