Serovar prevalence of Leptospira in semirural India and the development of an IgM-based indirect ELISA

  • Sara Chandy Pushpagiri Research Center, Pushpagiri Institute of Medical Sciences, Tiruvalla, India
  • Lokeshwaran Kirubanandhan Centre for BioSeparation Technology, (CBST) Vellore Institute of Technology, Vellore, India
  • Priya Hemavathy Christian Medical College, Vellore, India
  • Anees Mohammad Khadeeja Christian Medical College, Vellore, India
  • Siby Jacob Kurian Christian Medical College, Vellore, India
  • Krishnan Venkataraman Centre for BioSeparation Technology, (CBST) Vellore Institute of Technology, Vellore, India
  • Kristine Mørch National Center for Tropical Infectious Diseases, Haukeland University Hospital, Bergen, Norway
  • Dilip Mathai Apollo Institute of Medical Sciences and Research (AIMSR), Hyderabad, India
  • Anand Manoharan Pushpagiri Research Center, Pushpagiri Institute of Medical Sciences, Tiruvalla, India
Keywords: Leptospira serovars, recombinant LipL32, India

Abstract

Introduction: Leptospirosis is a major public health problem in India. However, it has been underreported and under-diagnosed due to a lack of awareness of the disease, a functional surveillance system, and appropriate laboratory diagnostic facilities.

Methodology: This multicenter study aimed to understand the Leptospira serovars causing leptospirosis in seven secondary-level hospitals in six states in India. Since early and accurate diagnosis of leptospirosis is one of the challenges faced by clinicians in India due to the poor specificity and sensitivity of commercially available diagnostic systems, an in-house indirect enzyme-linked immunosorbent assay (ELISA) was developed. Genomic DNA from L. interrogans serovar Canicola was used for polymerase chain reaction amplification, cloning, and expression of the lipL32 gene in E. coli to amplify, clone, and express the lipL32 gene.

Results: Australis was the common serovar seen at all the study centers. Serovar Icterohaemorrhagiae was seen in samples from Tamil Nadu and Assam. In-house ELISA was standardized using the purified recombinant LipL32 polypeptide and was used to evaluate serum. Subsequently, acute serum samples from leptospirosis patients (n = 60) were screened. Compared to the gold standard, the microscopic agglutination test, sensitivity and specificity of the in-house ELISA was 95% and 90%, respectively.

Conclusions: Understanding Leptospira serovars circulating in leptospirosis-endemic areas will help to formulate better vaccines. LipL32-based ELISA may serve as a valuable tool for early diagnosis of leptospirosis.

Author Biographies

Sara Chandy, Pushpagiri Research Center, Pushpagiri Institute of Medical Sciences, Tiruvalla, India

Scientist and Virology In Charge

Pushpagiri Research Center

Lokeshwaran Kirubanandhan, Centre for BioSeparation Technology, (CBST) Vellore Institute of Technology, Vellore, India

Research Scholar

Centre for BioSeparation Technology, (CBST) Vellore Institute of Technology, Vellore,India

Priya Hemavathy, Christian Medical College, Vellore, India

Research Scholar

Department of Gastroenterology

Christian Medical College

Vellore

Anees Mohammad Khadeeja, Christian Medical College, Vellore, India

Research Scholar

IDTRC

Christian Medical College

Vellore

Siby Jacob Kurian, Christian Medical College, Vellore, India

Research Scholar

IDTRC

Christian Medical College

Vellore

Krishnan Venkataraman, Centre for BioSeparation Technology, (CBST) Vellore Institute of Technology, Vellore, India

 

Professor

Centre for BioSeparation Technology, (CBST) Vellore Institute of Technology, Vellore,India

 

Kristine Mørch, National Center for Tropical Infectious Diseases, Haukeland University Hospital, Bergen, Norway

 

Professor

National Center for Tropical Infectious Diseases, Haukeland University Hospital, Bergen, Norway

Dilip Mathai, Apollo Institute of Medical Sciences and Research (AIMSR), Hyderabad, India

Dean

Apollo Institute of Medical Sciences and Research (AIMSR), Hyderabad,India

Anand Manoharan, Pushpagiri Research Center, Pushpagiri Institute of Medical Sciences, Tiruvalla, India

Head of Research and Development

Pushpagiri Research Center, Pushpagiri Institute of Medical Sciences, Tiruvalla, 689101,Ind

Published
2017-03-31
How to Cite
1.
Chandy S, Kirubanandhan L, Hemavathy P, Khadeeja AM, Kurian SJ, Venkataraman K, Mørch K, Mathai D, Manoharan A (2017) Serovar prevalence of Leptospira in semirural India and the development of an IgM-based indirect ELISA. J Infect Dev Ctries 11:234-241. doi: 10.3855/jidc.8067
Section
Original Articles