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. 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.
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. 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.
Entities:
Keywords:
16S rRNA; LipL32; leptospirosis; mPCR; nucleic acid amplification test
Authors: Ajay R Bharti; Jarlath E Nally; Jessica N Ricaldi; Michael A Matthias; Monica M Diaz; Michael A Lovett; Paul N Levett; Robert H Gilman; Michael R Willig; Eduardo Gotuzzo; Joseph M Vinetz Journal: Lancet Infect Dis Date: 2003-12 Impact factor: 25.071
Authors: Kanti Laras; Bao Van Cao; Khanthong Bounlu; Thi Kim Tien Nguyen; James G Olson; Sisouk Thongchanh; Nguyen Van Anh Tran; Kim Loan Hoang; Narain Punjabi; Ba Khiem Ha; Sam An Ung; Sithat Insisiengmay; Douglas M Watts; H James Beecham; Andrew L Corwin Journal: Am J Trop Med Hyg Date: 2002-09 Impact factor: 2.345
Authors: Marluce A Assunção Oliveira; Otávia L Caballero; Annamaria R Vago; Rudy A Harskeerl; Álvaro J Romanha; Sérgio D J Pena; Andrew J G Simpson; Matilde Cota Koury Journal: J Med Microbiol Date: 2003-02 Impact factor: 2.472
Authors: Jeremiah N Ngugi; Eric M Fèvre; Georgies F Mgode; Mark Obonyo; Ginethon G Mhamphi; Christina A Otieno; Elizabeth Anne Jessie Cook Journal: BMC Vet Res Date: 2019-11-08 Impact factor: 2.741
Authors: James P Herrera; Natalie R Wickenkamp; Magali Turpin; Fiona Baudino; Pablo Tortosa; Steven M Goodman; Voahangy Soarimalala; Tamby Nasaina Ranaivoson; Charles L Nunn Journal: PLoS Negl Trop Dis Date: 2020-12-31
Authors: Sabri A Rahman; Kuan H Khor; Siti Khairani-Bejo; Seng F Lau; Mazlina Mazlan; Azri Roslan; Soon H Goh Journal: J Vet Diagn Invest Date: 2021-06-19 Impact factor: 1.569
Authors: Andrei E Samoilov; Nataliya A Stoyanova; Nikolai K Tokarevich; Birgitta Evengard; Elena V Zueva; Yulia A Panferova; Yulia V Ostankova; Elena B Zueva; Diana E Valutite; Evgeniy V Kovalev; Anna R Litovko; Alexey U Goncharov; Alexandr V Semenov; Kamil Khafizov; Vladimir G Dedkov Journal: Int J Environ Res Public Health Date: 2020-06-14 Impact factor: 3.390