Molecular diagnostics for human leptospirosis

Abstract

Purpose of Review The definitive diagnosis of leptospirosis, which results from infection with spirochetes of the genus Leptospira, currently relies on the use of culture, serological testing (microscopic agglutination testing, MAT), and molecular detection. The purpose of this review is to describe new molecular diagnostics for Leptospira and discuss advancements in the use of available methods. Recent Findings Efforts have been focused on improving the clinical sensitivity of Leptospira detection using molecular methods. In this review, we describe a re-optimized pathogenic species-specific real-time PCR (targeting lipL32) that has demonstrated improved sensitivity; findings by two groups that real-time RT-PCRs targeting the 16S rrs gene can improve detection; and two new loop-mediated amplification techniques. Quantitation of leptospiremia, detection in different specimen types, and the complementary roles played by molecular detection and MAT will be discussed. Finally, a protocol for Leptospira strain subtyping using variable number tandem repeat targets and high-resolution melting will be described. Summary Molecular diagnostics have an established role for the diagnosis of leptospirosis and provide an actionable diagnosis in the acute setting. The use of real-time RT-PCR for testing serum/plasma and cerebrospinal fluid, when available, may improve the detection of Leptospira without decreasing clinical specificity.

The Association between Hydro-Meteorological Events and Leptospirosis Hospitalizations in Santa Catarina, Brazil

“…The maximum rates of hospitalization for Leptospirosis were identified in the months and/or subsequent month in which there were the highest number of current HE events, which is in fact related to Leptospirosis incubation period, which varies from 2 to 30 days, with an average of 7-10 days. [41] Moreover, the coastal macro-regions of health present the highest rates of hospitalization for this disease which is in agreement with the findings in other studies [36]. The regions that presented positive and significant correlations are mostly located in the coastal zone of the state of SC, which in recent years presented a significant number of hydrological events.…”

Evaluation of the sick returned traveler

“…Realtime PCR appears promising for leptospirosis, which has been historically challenging to confirm with culture and serology. 32 Increasingly, syndromic-based molecular panels that test for multiple pathogens are available, 33 raising hope that the diagnostic approach for returned sick travelers might be simplified and expedited. Multiplex PCR platforms for multiple bacterial, viral and protozoal causes of gastroenteritis are available and potentially useful for providing a rapid etiologic diagnosis in the travelers with diarrhea.…”

Leptospirosis in Poland in the years 2014-2017 – characteristics of infections and epidemiological surveillance data

“…This test could be done using the patient's blood, urine and tissues. Sequences of the most often applied primers in this molecular method is complementary to the genes: lipL32, lfb1 (only present in pathogenic species) or characteristic for the whole genotype: rrs (16S rRNA), secY, gyrB (genes of basic metabolism) (30,31). The DNA of spirochetes Leptospira spp.…”

“…Badanie to wykonuje się z krwi, moczu i tkanek chorego. Starterami najczęściej stosowanymi są sekwencje komplementarne do genów: lipL32, lfb1 (występują tylko u patogennych gatunków) lub charakterystycznych dla całego gatunku genów: rrs (16S rRNA), secY, gyrB (geny metabolizmu podstawowego) (30,31). DNA krętków Leptospira spp.…”