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Malaysian Journal of Medical Sciences
School of Medical Sciences, Universiti Sains Malaysia
ISSN: 1394-195X
Vol. 19, No. 3, 2012, pp. 9-16
Bioline Code: mj12030
Full paper language: English
Document type: Research Article
Document available free of charge

Malaysian Journal of Medical Sciences, Vol. 19, No. 3, 2012, pp. 9-16

 en Rapid Diagnosis of Leptospirosis by Multiplex PCR
Ahmed , Siti Aminah; Sandai, Doblin Anak; Musa , Suzana; Hoe , Chee Hock; Riadzi, Mehdi; Lau, Kwok Leong & Tang, Thean Hock

Abstract

Background: Traditionally, the most common diagnostic approach used for diagnosing leptospirosis was the demonstration of immune-seroconversion in acute and convalescent patient serum samples. Recently, a variety of molecular techniques, including conventional and real-time polymerase chain reaction (PCR), have been developed for the specific detection of pathogenic bacteria from the genus Leptospira check for this species in other resources . PCR is a sensitive, specific, and rapid technique that has been successfully used to detect several microorganisms; including those of clinical significance.
Methods: In this study, we developed a multiplex PCR (mPCR) assay for detecting Leptospira’s DNA. The mPCR assay detects both the 16S rRNA gene and the major outer membrane lipoprotein gene, which is known as LipL32. Representative serovars were tested from 10 species of Leptospira and 23 other species of bacteria.
Results: A positive result was obtained from all leptospiral serovars. The amplification sensitivity for the multiplex assay was 21.8 pg and 1 × 103 leptospires/ml. This mPCR assay has the potential to facilitate a rapid and sensitive diagnosis for acute leptospirosis.
Conclusion: The mPCR assay developed in this study can be used for the early detection of leptospirosis. The LipL32 gene could also serve as another target to aid in the efficient detection of leptospiral infection because using 2 sets of primers in mPCR increases the sensitivity and specificity of the test.

Keywords
16S rRNA, leptospirosis, LipL32, mPCR, nucleic acid amplification test

 
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Alternative site location: http://www.medic.usm.my/publication/mjms/

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