Establishment of Loop-mediated Isothermal Amplification Method for Rapid Detection of Haemophilus parasuis
Journal Title: 河南科技大学学报（自然科学版） - Year 2017, Vol 38, Issue 6
Abstract
A loop-mediated isothermal amplification ( LAMP) method was established for the simple, rapid, sensitive and specific detection of Haemophilus parasuis ( Hps) . Toking the 16S rRNA fragment of Hps as the targeting sequence, six specific gene regions were selected, and four specific primes ( c ( F3) 、c ( B3) 、c ( FIP) and c ( BIP) ) were designed in these regions. The LAMP rapid detection method for Hps was established by optimizing the reaction system, and detecting the sensitivity and specificity. The results show that the minimum detectable DNA quantity is 0. 241 pg/μL, when the primers c ( F3) : c ( FIP) or c ( B3) : c ( BIP) is 1 : 4, the concentration of Mg2+ is 2 mmol/L, and the concentration of d NTPs is 6 mmol/L. The sensitivity of the method is 100 times than that of polymerase chain reaction method. Furthermore, the method can only detect Hps, but can not detect Streptococcus, Salmonella, Escherichia coli, Bacillus subtilis, Actinobacillus pleuropneumoniae, Staphylococcus aureus.
Authors and Affiliations
Kui HOU, Ke DING, Shouye LIU, Zuhua YU, Zhanqin ZHAO
Keywords
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