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Leptospirosis in Febrile Patients: Diagnosis by serology and polymerase chain reaction

*, Khalid Perwez and *, Suman VB and *, Nayanatara Arun Kumar (2011) Leptospirosis in Febrile Patients: Diagnosis by serology and polymerase chain reaction. Journal of Biology and Lifescience, 2 (1: E3). pp. 1-9. ISSN 2157-6076

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Abstract

Introduction: Leptospirosis is a zoonotic disease found in all tropical and temperate areas of the world. Humans contract leptospira through mucosal or percutaneous exposure to environments contaminated by the urine of chronically infected animal sources mainly rodents,dogs, pigs and cattle. Objectives: It is known that IgM ELISA is used widely to diagnose leptospirosis in acute stage of illness. Our aim is to evaluate the sensitivity and specificity of a molecular technique like PCR in reference to the serological test Pan Bio, IgM ELISA, and its usefulness as a diagnostic test inthe diagnosis of leptospirosis. Methods: The patients included 85 males and 15 females, age 2yrs–78 yrs, (median age,40yrs). Common presenting symptoms were Fever, myalgia, jaundice, nausea and vomiting,oliguria and acute renal failure. Blood samples were drawn during the acute phase of illness,7.5 mean days from the onset of symptoms. All the samples were tested for IgM antibody by ELISA and for Leptospira DNA by PCR. Detection of antibody done by Pan Bio LeptospiraIgM ELISA. Detection of DNA by PCR: Extraction of DNA: was performed using the QIAGEN mini kit. Amplification of DNA: Primers used to amplify a 631 base-pair (bp) 5'-region of 16S rDNA of pathogenic leptospira, Detection of DNA: by Agarose gelelectrophoresis Results: All 100 samples were tested by both ELISA and PCR. 35 samples (35%) were positive for the presence of IgM antibodies by ELISA. PCR demonstrated DNA in 34 of the samples. A total of 28 samples were positive by both ELISA and PCR. The PCR demonstrated a sensitivity of 80 % and specificity of 90%. Conclusions: PCR is a sensitive and specific technique which can detect the presence of DNA in the very early stage of the disease; Results of PCR can give us a presumptive diagnosis of leptospirosis in early phase of illness (1-7days) compared to Pan Bio IgM ELISA. Hence PCR together with IgM ELISA can be used to confirm the diagnosis, in the acute stage (1-15days) of the infection.

Item Type: Article
Uncontrolled Keywords: Leptospirosis, IgM Antibody, ELISA, DNA, PCR
Subjects: Medicine > KMC Mangalore > Physiology
Depositing User: KMCMLR User
Date Deposited: 15 Nov 2012 09:56
Last Modified: 15 Nov 2012 09:56
URI: http://eprints.manipal.edu/id/eprint/77582

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