开发和验证一种用于检测猪肺炎支原体感染的 SIgA-ELISA 方法。
Development and validation of a SIgA-ELISA for the detection of Mycoplasma hyopneumoniae infection.
机构信息
Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
出版信息
Vet Microbiol. 2010 Jul 14;143(2-4):410-6. doi: 10.1016/j.vetmic.2009.11.038. Epub 2009 Dec 4.
An alternative indirect enzyme-linked immunosorbent assay (ELISA) for the detection of Mycoplasma hyopneumoniae secretory IgA (SIgA) antibody (SIgA-ELISA) was developed using an adhesin (P97R1) of M. hyopneumoniae produced in Escherichia coli. The SIgA-ELISA assay was validated by the comparison with a nested-PCR assay and a commercial M. hyopneumoniae antibody detection kit (IgG-ELISA). Two hundred and sixty nasal swab samples, bronchoalveolar lavage fluids or serum samples were prepared for SIgA-ELISA validation from a M. hyopneumoniae-free farm, a M. hyopneumoniae vaccinated farm and two M. hyopneumoniae contaminated farms. The results showed that the SIgA-ELISA assay could distinguish the M. hyopneumoniae infection from M. hyopneumoniae vaccinated pigs, which was impossible for the current commercial M. hyopneumoniae antibody detection kits. The diagnostic sensitivity (DSN), specificity (DSP) and accuracy of the SIgA-ELISA were 97.0%, 94.4% and 95.8%, respectively and were compared with nested-PCR on 260 field nasal swab samples. The results of repeatability tests revealed that the coefficients of variation of swab samples within and between runs were less than 10%. This SIgA-ELISA is a needle-free detection methodology for large-scale surveys of M. hyopneumoniae infection.
一种用于检测猪肺炎支原体分泌型 IgA（SIgA）抗体的间接酶联免疫吸附试验（ELISA）的替代方法被开发出来，该方法使用在大肠杆菌中产生的猪肺炎支原体黏附素（P97R1）。通过与巢式 PCR 检测方法和商业性猪肺炎支原体抗体检测试剂盒（IgG-ELISA）的比较，对 SIgA-ELISA 检测方法进行了验证。从一个无猪肺炎支原体的农场、一个接种过猪肺炎支原体疫苗的农场和两个受猪肺炎支原体污染的农场中，共制备了 260 份鼻拭子样本、支气管肺泡灌洗液或血清样本，用于 SIgA-ELISA 验证。结果表明，SIgA-ELISA 检测方法可以区分猪肺炎支原体感染和接种过猪肺炎支原体疫苗的猪，而目前的商业性猪肺炎支原体抗体检测试剂盒则无法做到这一点。在对 260 份现场鼻拭子样本进行检测时，SIgA-ELISA 的诊断敏感性（DSN）、特异性（DSP）和准确性分别为 97.0%、94.4%和 95.8%，与巢式 PCR 检测方法相比。重复性试验结果表明，在同一批次和不同批次运行中，拭子样本的变异系数均小于 10%。这种 SIgA-ELISA 是一种无针检测方法，可用于大规模调查猪肺炎支原体感染。
Zotero 插件