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| The Application of Hydroxy Naphthol Blue in Loop-mediated Isothermal Amplification of Pseudorabies virus |
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Abstract The traditional diagnostic method for Pseudorabies virus(PRV) has the defect of cumbersome procedures, long period, high cost and poor specificity. Thus we built a sensitive and specific loop-mediated isothermal amplification(LAMP) method for the detection of PRV, and used hydroxy naphthol blue(HNB) as indicator to judge the reaction result. The highly conserved gB (glycoprotein B)gene was selected to design primers. The detection could be finished within 45 min after the reaction system was established. Both LAMP and PCR were highly specific to PRV，but he LAMP-based assay, whose detection limit was 102 copies of the target sequence，was 10 times more sensitive than that of the PCR assay. In HNB test, when the ultimate concentration of HNB was 150 μmol/L, the color contrast of the LAMP products between positive samples and negative control was obvious and the sensisity of LAMP reaction was the best.The detection of clinical samples showed that the LAMP reaction had the similar detection rate with the PCR assay. In this experiment, the LAMP method is a highly specific and sensitive detection technology, which can be used instead of traditional PCR method.
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Received: 09 June 2011
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NI He-Jia, CHENG Xue, WANG Jing, GAO Ze-Ping, WEI Pan-Pan, TAN Jia-Li, SUN Jing, LI Hai-Tao, GAO Ji-Guo. Isolation and Screening of High Virulent Bacillus thuringiensis Strains Against Lepidopteran Pests in Some Areas of Liaoning Province[J]. 农业生物技术学报, 2019, 27(8): 1513-1520. |
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