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鱼类诺卡氏菌病(fish nocardiosis)是一种常见的严重危害全球水产养殖业的鱼类疾病。鰤鱼诺卡氏菌(Nocardia seriolae)是鱼类诺卡氏菌病的主要病原菌之一。本研究对已报道的鰤鱼诺卡氏菌17个不同菌株全基因组数据进行比对分析，筛选获得种内保守、种间变异的特异性基因组片段，并设计特异性引物，建立鰤鱼诺卡氏菌特异性PCR检测方法。设计的特异性引物仅能扩增获得鰤鱼诺卡氏菌的特异性片段，其他近缘菌和常见菌均无PCR扩增片段，且PCR检测灵敏度核酸浓度达80 pg/μL，菌液浓度达8.6 CFU/μL。用该PCR检测方法检测人工感染鰤鱼诺卡氏菌和阴性对照组杂交鳢各组织，结果显示，只能从感染鰤鱼诺卡氏菌的鱼组织中扩增出阳性片段。本研究基于鰤鱼诺卡氏菌不同菌株全基因组数据比对分析，所建立的PCR检测方法比之前的分子检测方法更具科学性、可靠性。该PCR检测方法可以提供一种快速、准确、灵敏的鰤鱼诺卡氏菌检测方式。
Abstract：Fish nocardiosis is a common fish disease that seriously harms the global aquaculture industry. Nocardia seriolae is one of the main pathogens of fish nocardiosis. In this study, the specific gene fragments with intraspecific conservation and interspecific variation were screened by comparing and analyzing the whole genomes of 17 reported N. seriolae strains, and specific primers were designed to establish a specific PCR method for the detection of N. seriolae. The designed primers can only amplify the specific fragments of N. seriolae, no PCR fragments showed in other bacteria, and the sensitivity of PCR detection is up to 80 pg/μL, 8.6 CFU/μL. The PCR method was used to detect the tissues of hybrid snakehead with N. seriolae infection or negative control, the results showed that the positive fragments could only be amplified from the tissues of N. seriolae-infected fish. In this study, based on the comparison and analysis of the whole genome data of different strains of N. seriolae, the established PCR method is more reasonable and reliable than the previous molecular detection method. This PCR method can provide a rapid, accurate and sensitive detection for N. seriolae.
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基本信息:
DOI：10.13417/j.gab.040.002656
中图分类号:S943
引用信息:
[1]陈国权,黄碧艳,张子雯,等.鰤鱼诺卡氏菌特异性PCR检测方法的建立[J].基因组学与应用生物学,2021,40(Z2):2656-2665.DOI:10.13417/j.gab.040.002656.
基金信息:
广东海洋大学“冲一流”学科建设科研项目(231419017); 深圳大鹏新区产业发展专项(KJYF202001-08); 深圳市科技计划项目基础研究(自由探索)项目(JCYJ20180306173022502; JCYJ20180507183240459)共同资助
2020-09-25
2020-09-25
2020-09-25