Centre for Research and Development of Biomedical and Basic Health Technology, National Institute of Health Research and Development, Ministry of Health, Jakarta, Indonesia.
Centre for Research and Development of Biomedical and Basic Health Technology, National Institute of Health Research and Development, Ministry of Health, Jakarta, Indonesia.
J Microbiol Methods. 2021 May;184:106198. doi: 10.1016/j.mimet.2021.106198. Epub 2021 Mar 10.
In diphtheria laboratory examinations, the PCR test can be applied to isolates and clinical specimens. This study aimed to develop a PCR assay to identify the species and toxigenicity of diphtheria-causing bacteria, including the prediction of some NTTB types. Seven reference isolates, four synthetic DNA samples, 36 stored isolates, and 487 clinical samples used for PCR optimization. The PCR results was confirmed by DNA sequence analysis. The results of the PCR examination of the 7 reference isolates and 36 stored isolates were similar to the results obtained using conventional methods as gold standard, both for diphtheria-causing and non-diphtheria-causing bacteria. The validation of the PCR results using DNA sequence analysis showed that there was no mispriming or misamplification. The multiplex PCR assay developed in this study could correctly identify the species and toxigenicity of diphtheria-causing bacteria, including the prediction of some NTTB types not yet covered by established PCR methods.
在白喉实验室检查中，可以将 PCR 检测应用于分离株和临床标本。本研究旨在开发一种 PCR 检测方法，以鉴定引起白喉的细菌的种属和毒力，包括对一些 NTTB 型的预测。该研究使用了 7 个参考分离株、4 个合成 DNA 样本、36 个储存分离株和 487 个临床样本进行 PCR 优化。PCR 结果通过 DNA 序列分析进行确认。7 个参考分离株和 36 个储存分离株的 PCR 检测结果与传统方法（金标准）相似，无论是对白喉菌还是非白喉菌的检测结果都相似。使用 DNA 序列分析对 PCR 结果进行验证表明，不存在引物错配或非特异性扩增。本研究中开发的多重 PCR 检测方法可以正确鉴定引起白喉的细菌的种属和毒力，包括对一些尚未被现有 PCR 方法涵盖的 NTTB 型的预测。
