Diversity and evolution of methods and practices for the molecular diagnosis of congenital toxoplasmosis in France: a 4-year survey

Abstract

The prenatal diagnosis of congenital toxoplasmosis is currently based upon molecular biology using a sample of amniotic fluid. The vast majority of centres globally (and all centres in France) performing this diagnosis use 'in house' or laboratory-developed PCR assays. This may be the source of considerable inter-laboratory variation in the performances of the assays, hampering any valuable comparison of data among different centres. The present study was based upon questionnaires that were sent to 21-25 centres between 2002 and 2005 enquiring about methods and practices of the PCR-based prenatal diagnosis of congenital toxoplasmosis. An extreme diversity of PCR methods and practices was observed. Thus, in 2005, 35 PCR methods, differing in one of the main steps of the whole process, were reported as being in use for routine diagnosis, with nine centres using two or three methods. We provide comprehensive information on the extraction methods, DNA targets, primer pairs and detection methods used for this diagnosis, as well as their evolution, during the period of study. Interestingly, in this period (2002-2005), a rapid progression of the number of laboratories using real-time PCR technology, which increased from four to 19, was observed. We also studied general PCR practices concerning, for example, the number of reaction tubes used for each biological sample and the inclusion of controls. The return of information in a yearly report provided the opportunity for writing proposals aiming to improve laboratory practices for this diagnosis at the national level. The high diversity of methods and practices currently used emphasizes the need for external quality assessment of the performances of the molecular diagnostic methods.