title: Rapid diagnosis of leptospirosis by multiplex PCR.
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. 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 × 10(3) leptospires/ml. This mPCR assay has the potential to facilitate a rapid and sensitive diagnosis for acute leptospirosis.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Advanced Medical & Dental Institute, Universiti Sains Malaysia, 11800, Penang, Malaysia.
authors.forename: Siti Aminah
authors.initials: SA
authors.lastname: Ahmed
authors.forename: Doblin Anak
authors.initials: DA
authors.lastname: Sandai
authors.forename: Suzana
authors.initials: S
authors.lastname: Musa
authors.forename: Chee Hock
authors.initials: CH
authors.lastname: Hoe
authors.forename: Mehdi
authors.initials: M
authors.lastname: Riadzi
authors.forename: Kwok Leong
authors.initials: KL
authors.lastname: Lau
authors.forename: Thean Hock
authors.initials: TH
authors.lastname: Tang
keywords: 16S rRNA; LipL32; leptospirosis; mPCR; nucleic acid amplification test