Open AccessFollowing the course of human leptospirosis: evidence of a critical threshold for the vital prognosis using a quantitative PCR assay
Author(s) -
Truccolo Johann,
Serais Ophélie,
Merien Fabrice,
Perolat Philippe
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10904.x
Subject(s) - leptospirosis , leptospira , biology , detection limit , microbiology and biotechnology , virology , chromatography , chemistry
In order to follow the course of acute human leptospirosis, an ELISA microtiter plate hybridization method was developed for the quantitative determination of Leptospira spp. in biological samples after PCR. The biotin‐labelled amplified product (331 bp from the rrs gene) was hybridized with a complementary capture probe covalently linked onto aminated polystyrene wells, and detected using a colorimetric reaction. The mean detection limit was 50 copies per 10 μl. In a prospective study of human leptospirosis cases, we obtained evidence that a density of 10 4 leptospires per ml of blood is a critical threshold for the vital prognosis of the patients. The practicability of the method makes it suitable for use in tropical areas for multicentric studies. Such studies could lead to a better knowledge of the natural history of the human disease. The method is also suitable for experimental evaluation of improved antibiotic treatments for leptospirosis.