Gou Hongchao, Li Juan, Cai Rujian, Song Shuai, Li Miao, Yang Dongxia, Jiang Zhiyong, Li Yan, Chu Pinpin, Li Chunling
Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China; Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China; Guangdong Open Laboratory of Veterinary Public Health, Guangzhou, China.
Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China; Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China; Guangdong Open Laboratory of Veterinary Public Health, Guangzhou, China.
J Microbiol Methods. 2018 Jan;144:67-72. doi: 10.1016/j.mimet.2017.11.005. Epub 2017 Nov 9.
Haemophilus parasuis infection is of considerable economic importance in the swine industry due to high morbidity and mortality in naive swine populations. Accurate detection and identification of the causative agent are difficult, yet necessary, for disease control. In this study, a simple and rapid method of cross-priming amplification (CPA) with a vertical flow (VF) visualization strip was established to detect H. parasuis. The reaction can specifically identify 15 serovar reference strains and 57 clinically isolated strains of H. parasuis, with a detection limit of 14CFU. The performance of the CPA-VF assay was evaluated and compared with that of species-specific PCR by testing 62 clinical culture-positive specimens of H. parasuis. The entire process, from specimen processing to analysis of the results, can be completed in 2h without a complicated apparatus. The convenience and speed of the CPA-VF assay in this study make it a suitable choice for epidemiological investigation and point-of-care testing (POCT) for H. parasuis infection.
由于初生猪群中发病率和死亡率较高，副猪嗜血杆菌感染在养猪业中具有相当重要的经济意义。准确检测和鉴定病原体对于疾病控制而言虽困难但却必要。在本研究中，建立了一种采用垂直流（VF）可视化条带的交叉引物扩增（CPA）简单快速方法来检测副猪嗜血杆菌。该反应能够特异性鉴定15株副猪嗜血杆菌血清型参考菌株和57株临床分离菌株，检测限为14CFU。通过检测62份副猪嗜血杆菌临床培养阳性标本，对CPA-VF检测方法的性能进行了评估，并与种特异性PCR进行了比较。从标本处理到结果分析的整个过程无需复杂仪器，2小时即可完成。本研究中CPA-VF检测方法的便利性和速度使其成为副猪嗜血杆菌感染流行病学调查和即时检测（POCT）的合适选择。
