Yang X, Qi M W, Zhang Z Z, Gao C, Wang C Q, Lei W Q, Tan L, Zhao J L, Fang R, Hu M
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, PR China.
J Parasitol. 2017 Apr;103(2):161-167. doi: 10.1645/16-157. Epub 2017 Jan 18.
Haemonchus contortus is one of the most significant strongylid nematodes infecting small ruminants, and it causes great economic losses to the livestock industry worldwide. Accurate diagnosis of H. contortus is crucial to control strategies. Traditional microscopic examinations are the most common methods for the diagnosis of H. contortus , but they are time-consuming and inaccurate. Molecular methods based on PCR are more accurate, but need expensive machines usually only used in the laboratory. Loop-mediated isothermal amplification (LAMP) is a rapid, simple, specific, and sensitive method that has been widely used to detect viruses, bacteria, and parasites. In the present study, a LAMP method targeting ribosomal ITS-2 gene for detection of the H. contortus in goat fecal samples has been established. The established LAMP method was H. contortus specific, and the sensitivity of LAMP was the same as that of the H. contortus species-specific PCR, with the lowest DNA level detected as being 1 pg. Examination of the clinical samples indicated that the positive rate of LAMP was higher than that of PCR, but no statistical difference was observed between LAMP and PCR (χ = 17.991, P = 0.053). In conclusion, a LAMP assay with a high specificity and a good sensitivity has been developed to detect H. contortus infection in goats. The established LAMP assay is useful for clinical diagnosis of H. contortus .
捻转血矛线虫是感染小反刍动物的最重要的圆线虫之一,它给全球畜牧业造成了巨大的经济损失。准确诊断捻转血矛线虫对于防控策略至关重要。传统的显微镜检查是诊断捻转血矛线虫最常用的方法,但它们耗时且不准确。基于聚合酶链反应(PCR)的分子方法更准确,但需要通常仅在实验室使用的昂贵仪器。环介导等温扩增技术(LAMP)是一种快速、简便、特异且灵敏的方法,已被广泛用于检测病毒、细菌和寄生虫。在本研究中,已建立了一种针对核糖体ITS-2基因的LAMP方法,用于检测山羊粪便样本中的捻转血矛线虫。所建立的LAMP方法对捻转血矛线虫具有特异性,LAMP的灵敏度与捻转血矛线虫种特异性PCR相同,检测到的最低DNA水平为1皮克。对临床样本的检测表明,LAMP的阳性率高于PCR,但LAMP与PCR之间未观察到统计学差异(χ = 17.991,P = 0.053)。总之,已开发出一种具有高特异性和良好灵敏度的LAMP检测方法,用于检测山羊的捻转血矛线虫感染。所建立的LAMP检测方法对捻转血矛线虫的临床诊断有用。