基于聚合酶链式反应（PCR）检测鱼类病原菌杀鲑气单胞菌。II. 对感染鱼类的三组PCR引物进行进一步评估与验证

PCR-based assays for the fish pathogen Aeromonas salmonicida. II. Further evaluation and validation of three PCR primer sets with infected fish.

作者信息

Byers Helen K, Cipriano Rocco C, Gudkovs Nicholas, Cranel Mark St J

机构信息

Australian Animal Health Laboratory, CSIRO Livestock Industries, Geelong, VIC.

出版信息

Dis Aquat Organ. 2002 May 10;49(2):139-44. doi: 10.3354/dao049139.

DOI:10.3354/dao049139

PMID:12078982

Abstract

Two Aeromonas salmonicida-specific polymerase chain reaction (PCR) tests and 1 A. salmonicida subsp. salmonicida-specific PCR test were used to screen salmonid populations that were either overtly or covertly infected with A. salmonicida subsp. salmonicida. It was demonstrated that these PCR assays could be used to replace the biochemical testing currently employed to confirm the identity of A. salmonicida isolates cultured from infected fish. The AP and PAAS PCR assays were also capable of direct detection of A. salmonicida in overtly infected fish, with mucus, gill and kidney samples most likely to yield a positive result. Culture was a more reliable method for the direct detection of A. salmonicida in covertly infected salmonids than was the direct PCR testing of tissue samples, with the AP and PAAS PCRs having a lower detection limit (LDL) of approximately 4 x 10(5) colony-forming units (CFU) g(-1) sample.

摘要

使用两种杀鲑气单胞菌特异性聚合酶链反应（PCR）检测和1种杀鲑气单胞菌杀鲑亚种特异性PCR检测，对明显或隐匿感染杀鲑气单胞菌杀鲑亚种的鲑科鱼类种群进行筛查。结果表明，这些PCR检测方法可用于替代目前用于确认从感染鱼类中培养出的杀鲑气单胞菌分离株身份的生化检测。AP和PAAS PCR检测还能够直接检测明显感染的鱼类中的杀鲑气单胞菌，黏液、鳃和肾脏样本最有可能得出阳性结果。对于隐匿感染的鲑科鱼类，培养法比组织样本的直接PCR检测法更可靠地直接检测杀鲑气单胞菌，AP和PAAS PCR的检测下限（LDL）约为4×10⁵菌落形成单位（CFU）g⁻¹样本。