Leptospirosis in Febrile Patients: Diagnosis by Serology and Polymerase Chain Reaction

Leptospirosis in Febrile Patients: Diagnosis by Serology and Polymerase Chain Reaction

by Khalid Perwez, Suman VB, Nayanatara Kumar, Mohammad Hussain
     
 

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… See more details below

Overview

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 in
the 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 Leptospira
IgM 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 gel
electrophoresis
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.

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Product Details

ISBN-13:
2940148404033
Publisher:
ReadCycle
Publication date:
09/16/2013
Sold by:
Barnes & Noble
Format:
NOOK Book
File size:
0 MB

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