Multiplex PCR on leptospiral isolates from Kolenchery, Kerala, India.

Human leptospirosis causes severe multiorgan dysfunctions that may end in multiorgan failure and death. The methods in hand for diagnosis of leptospirosis like culture, ELISA and MAT only help to confirm the disease, and are of little value in early detection. The aim of this study was to find out if the two sets of primers described earlier could detect all the isolates from the area, for the purpose of using the resultant database for early detection of leptospires in future from clinical specimens. The study was done on culture isolates from Jan 2000 to June 2002 attending the department of medicine, MOSC medical college hospital, Kolenchery, Kerala, India. DNA of 45 culture isolates were amplified by multiplex PCR using two sets of previously described primers, G1, G2 and B 64-I, B 64-II. Specific amplifications of either 285 or 563 bp size were obtained from all isolates included in the study indicating the utility of the multiplex PCR in the rapid detection of leptospires in clinical samples.

Lateral-flow assay for rapid serodiagnosis of human leptospirosis.

An assay device for the rapid detection of Leptospira-specific immunoglobulin M (IgM) antibodies in human sera is presented. The sensitivity (85.8%) and specificity (93.6%) of the assay compared well (91.9% agreement) with those of an IgM enzyme-linked immunosorbent assay routinely used in the serodiagnosis of leptospirosis. The sensitivity of the assay varied with the stage of the disease. The assay uses stabilized components and is simply performed by the addition of serum and sample fluid to the sample well of the assay device. The assay is read after 10 min, and a positive result is obtained when staining of the test line is observed.

Latex based, rapid and easy assay for human leptospirosis in a single test format.

Leptospirosis is an often severe disease which requires prompt treatment. Laboratory testing is required to reach a valid diagnosis. An agglutination assay for the detection of Leptospira-specific antibodies consisting of individually wrapped agglutination cards containing a stable, dried detection reagent is evaluated. The assay is simply performed by suspending the dried reagent with a drop of serum. The result is obtained within 30 s. The sensitivity of the assay varied with the stage of the disease and was 72.3% for samples collected during the first 10 days of the illness and 88.2% for samples collected at a later stage. The specificity was 93.9% and 89.8%, respectively. These characteristics make the test ideal for use in areas where the disease is common and where laboratory support is not routinely available.