MAGNETIC IMMUNO CAPTURE PCR ASSAY (MIPA) - DETECTION OF LEPTOSPIRA-BORGPETERSENII SEROVAR HARDJO

Magnetic immune PCR assay (MIPA) was developed for the rapid detection of leptospires excreted in urine samples (n = 59) collected from 35 e xperimentally infected cattle. The immuno-magnetic separation of lepto spires from inhibitors in frozen formalin fixed bovine urine prior to PCR detection resulted in a marked improvement on previous detection m ethods. MIPA is a rapid 5 step protocol requiring 70 mins preparation time prior to amplification, which consistently detects 10(1) organism s. MIPA detected 76% (38/50) of culture positive urines and in additio n three urines that were culture negative were shown to be positive by this method of detection. Consequently we conclude that whilst MIPA i s an improvement on previously published PCR detection methods, the cu lture of the organism is still the standard against which other detect ion methods have to be compared. (C) 1997 Elsevier Science B.V.