Comparative analysis of current diagnostic PCR assays in detecting pathogenic Leptospira isolates from environmental samples
Journal Title: Asian Pacific Journal of Tropical Medicine - Year 2019, Vol 12, Issue 10
Abstract
Objective: To compare the efficiency of routine diagnostic PCR assays in detecting pathogenic Leptospira isolated from water and soils. Methods: Seven routine assays targeting six genes (lipL32, flaB, gyrB, lfb1, secY and ligB) were evaluated and compared on the cultures of two groups of pathogenic Leptospira from different sources. One group included 19 described reference strains recovered from infected human or animals, and another group included 22 environmental isolates from recreational and residential sites in Malaysia. The latter have been confirmed for presence of pathogenic Leptospira DNA. PCR positivity or detection sensitivity of each assay was determined and compared between the two groups. Results: Validation on reference strains showed 100.0% PCR sensitivity for all assays except ligB-PCR (95.0%) that failed to amplify Leptospira interrogans serovar Pomona. In marked contrast, there was a notable decline in sensitivity in the environmental isolates (lipL32-PCR, 95.5%; flaB-PCR, 90.9%; gyrB-PCR, 77.3%; lfb1-PCR, 59.1%; secY-PCRs, 40.9% G1/G2- PCR, 36.4%; ligB-PCR, 13.6%), implying a large genetic distance between the two groups, as well as nucleotide polymorphism among environmental isolates. Conclusions: High proportion of false-negative PCR results suggests a need of prudent selection of primers in detecting environmental pathogenic Leptospira. These findings offer valuable insights on the extensive biodiversity of genus Leptospira and its impact on the efficacy and development of molecular detection tool.
Authors and Affiliations
Vasantha Kumari Neela
Keywords
Related Articles
- EP ID EP663625
- DOI 10.4103/1995-7645.269908
- Views 82
- Downloads 0
Calcium carbonate supplementation causes memory impairment in mice
Objective: To investigate the influence of calcium carbonate supplementation on cognitive function in mice. Methods: Mice were fed diets containing 1.0% calcium carbonate for 8 weeks, following which they were evaluated...
Herbal remedies, vaccines and drugs for dengue fever: Emerging prevention and treatment strategies
Dengue fever is a disease that is caused by five dengue virus (DENV) serotypes. It is endemic in more than 128 countries. Millions of people are affected by this disease. But still, there is no specific treatment avail...
Epidemiological status of malaria in Iran, 2011–2014
Objectives: To aim to determine the status and epidemiological aspects of malaria in Iran in favour of gaining a better understanding of the national control of malaria. Methods: This epidemiological study examined the s...
Forecasting the number of zoonotic cutaneous leishmaniasis cases in south of Fars province, Iran using seasonal ARIMA time series method
Objective: To predict the trend of cutaneous leishmaniasis and assess the relationship between the disease trend and weather variables in south of Fars province using Seasonal Autoregressive Integrated Moving Average (SA...
Zika virus infection and its emerging trends in Southeast Asia
Zika virus is a mosquito-borne flavivirus that represents a public health emergency at the ongoing epidemic. Previously, this rare virus was limited to sporadic cases in Africa and Asia until its emergence in Brazil, Sou...