Li Zhenpeng, Guan Hongxia, Wang Wei, Gao He, Feng Weihong, Li Jie, Diao Baowei, Zhao Hongqun, Kan Biao, Zhang Jingyun
State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Wuxi Center for Disease Control and Prevention, Wuxi, China.
Front Cell Infect Microbiol. 2021 Feb 25;11:639473. doi: 10.3389/fcimb.2021.639473. eCollection 2021.
and are common diarrheal pathogens of great public health concern. A multiplex TaqMan-based real-time PCR assay was developed on the BD MAX platform; this assay can simultaneously detect and differentiate and directly from human fecal specimens. The assay includes two reactions. One reaction, BDM-VC, targets the gene , the cholera toxin (CT) coding gene , the O1 serogroup specific gene , and the O139 serogroup specific gene of . The other, BDM-VP, targets the gene and the toxin coding genes and of . In addition, each reaction contains a sample process control. When evaluated with spiked stool samples, the detection limit of the BD MAX assay was 195-780 CFU/ml for and 46-184 CFU/ml for , and the amplification efficiency of all genes was between 95 and 115%. The assay showed 100% analytical specificity, using 63 isolates. The BD MAX assay was evaluated for its performance compared with conventional real-time PCR after automated DNA extraction steps, using 164 retrospective stool samples. The overall percent agreement between the BD MAX assay and real-time PCR was ≥ 98.8%; the positive percent agreement was 85.7% for , 100% for /, and lower (66.7%) for because of a false negative. This is the first report to evaluate the usage of the BD MAX open system in detection and differentiation of and directly from human samples.
[具体病原体名称1]和[具体病原体名称2]是引起腹泻的常见病原体，备受公共卫生关注。在BD MAX平台上开发了一种基于多重TaqMan的实时PCR检测方法；该检测方法可直接从人类粪便标本中同时检测和区分[具体病原体名称1]和[具体病原体名称2]。该检测方法包括两个反应。一个反应BDM-VC，靶向[具体病原体名称1]的[基因名称1]、霍乱毒素（CT）编码基因[基因名称2]、O1血清群特异性基因[基因名称3]和O139血清群特异性基因[基因名称4]。另一个反应BDM-VP，靶向[具体病原体名称2]的[基因名称5]以及毒素编码基因[基因名称6]和[基因名称7]。此外，每个反应都包含一个样本处理对照。在用加标的粪便样本进行评估时，BD MAX检测方法对[具体病原体名称1]的检测限为195 - 780 CFU/ml，对[具体病原体名称2]的检测限为46 - 184 CFU/ml，所有基因的扩增效率在95%至115%之间。使用63株分离株，该检测方法显示出100%的分析特异性。在自动DNA提取步骤后，使用164份回顾性粪便样本，将BD MAX检测方法与传统实时PCR的性能进行了评估。BD MAX检测方法与实时PCR之间的总体一致性百分比≥98.8%；[具体病原体名称1]的阳性一致性百分比为（此处原文有误，应为85.7%），[具体病原体名称1]/[具体病原体名称2]的阳性一致性百分比为100%，而[具体病原体名称[此处原文有误]的阳性一致性百分比较低（66.7%），因为存在假阴性。这是第一份评估BD MAX开放系统直接从人类样本中检测和区分[具体病原体名称1]和[具体病原体名称2]的使用情况的报告。
