Open AccessLoop‐mediated isothermal amplification for the rapid detection of Haemophilus parasuis
Author(s) -
Chen HaoTai,
Chu YueFeng,
Liu YongSheng,
Zhang Jie,
Lu ZhongXin
Publication year - 2010
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.2010.00728.x
Subject(s) - loop mediated isothermal amplification , haemophilus , biology , microbiology and biotechnology , detection limit , primer (cosmetics) , virology , bacteria , dna , chromatography , chemistry , genetics , organic chemistry
Haemophilus parasuis infection is of considerable economic importance in the swine industry due to the high costs associated with treatment and loss of animals all over the world. In the present study, loop‐mediated isothermal amplification (LAMP) is described for the rapid and specific detection of this species. A primer set derived from the inf B gene of H. parasuis was used to validate the assay using 15 H. parasuis reference strains, 39 clinical isolates, 75 positive samples, and 18 other pathogens. The results indicated that positive reactions were confirmed for all H. parasuis strains and specimens by LAMP after 45 min reaction at 65 °C in a water bath, and no cross‐reactivity was observed from other non‐ H. parasuis strains. The detection limit of the conventional PCR was 25 copies, while that of the LAMP was five copies per tube. Therefore, the sensitivity of LAMP was higher than that of PCR. LAMP is likely to be more suitable as a routine diagnostic tool, especially in clinics without complicated equipment such as thermal cycling machines and electrophoresis apparatus. In these scenarios, the H. parasuis LAMP assay has the potential for field diagnosis.