Thontiravong Aunyaratana, Tantilertcharoen Rachod, Tuanudom Ranida, Sreta Donreuthai, Thanawongnuwech Roongroje, Amonsin Alongkorn, Oraveerakul Kanisak, Kitikoon Pravina
Emerging and Re-emerging Infectious Diseases in Animals Research Unit, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
J Vet Diagn Invest. 2011 Sep;23(5):1017-21. doi: 10.1177/1040638711416620.
A recently emerged H1N1 Influenza A virus (pandemic H1N1 (pH1N1)) with a Swine influenza virus (SIV) genetic background spread globally from human-to-human causing the first influenza virus pandemic of the 21st century. In a short period, reverse zoonotic cases in pigs followed by a widespread of the virus in the pig population were documented. The implementation of effective control strategies, rapid diagnosis, and differentiation of such virus from endemically circulating SIV in the various swine populations of the world is needed. To address the problem, a multiplex reverse transcription polymerase chain reaction assay utilizing a combination of the PB1, H1, and N1 primers that can rapidly and simultaneously subtype and screen for the presence of pH1N1 virus infection in Thai pigs was developed. The assay had 100% specificity and did not amplify genetic material from other subtypes of SIV, seasonal H1N1 human influenza (huH1N1) virus, highly pathogenic influenza H5N1 virus, and other important swine respiratory viral pathogens. The assay was able to both detect and subtype pH1N1 virus as low as 0.1-50% tissue culture infective doses/ml (TCID(50)/ml). The assay was used to screen 175 clinical samples obtained from SIV suspected cases, of which 6 samples were pH1N1 positive and were confirmed through virus isolation and whole genome sequencing. The results of the study suggested that the assay would be useful for the rapid diagnosis of pH1N1 in suspected Thai swineherds, where genetics of the endemically circulating SIV differ from the strains circulating in North American and European herds.
一种最近出现的具有猪流感病毒（SIV）基因背景的甲型H1N1流感病毒（大流行H1N1（pH1N1））在全球范围内通过人际传播，引发了21世纪的首次流感病毒大流行。在短时间内，有记录显示猪出现了反向人畜共患病病例，随后该病毒在猪群中广泛传播。需要实施有效的控制策略、进行快速诊断，并将这种病毒与世界各猪群中地方性流行的SIV区分开来。为了解决这个问题，开发了一种多重逆转录聚合酶链反应检测方法，该方法使用PB1、H1和N1引物的组合，能够快速同时对泰国猪中的pH1N1病毒感染进行亚型鉴定和筛查。该检测方法具有100%的特异性，不会扩增来自其他SIV亚型、季节性H1N1人类流感（huH1N1）病毒、高致病性H5N1流感病毒以及其他重要猪呼吸道病毒病原体的遗传物质。该检测方法能够检测和鉴定低至0.1 - 50%组织培养感染剂量/毫升（TCID(50)/毫升）的pH1N1病毒。该检测方法用于筛查从疑似SIV病例中获得的175份临床样本，其中6份样本pH1N1呈阳性，并通过病毒分离和全基因组测序得到证实。研究结果表明，该检测方法对于疑似泰国猪群中pH1N1的快速诊断将是有用的，在这些猪群中，地方性流行的SIV的基因与北美和欧洲猪群中流行的毒株不同。
