A Novel Molecular Method for Simultaneous Identification of Vibrio parahaemolyticus 57 K-Serogroups Using Probe Melting Curve Analysis.
A Novel Molecular Method for Simultaneous Identification of Vibrio parahaemolyticus 57 K-Serogroups Using Probe Melting Curve Analysis.
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使用探针熔解曲线分析同时鉴定副溶血弧菌 57 K 血清群的新分子方法。
DOI：
10.3389/fcimb.2021.594808
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发表时间：
2021
影响因子：
5.7
通讯作者：
Hu Q
中科院分区：
文献类型：
--
作者：
Lu L;Li M;Li Y;Jiang M;Jiang Y;Shi X;Zuo L;Wang L;Bian S;Qiu Y;Cai R;Liao Y;Li Q;Li L;Hu Q
副溶血性弧菌的血清分型对于弧菌病感染暴发的监测和检测至关重要，已在许多国家得到广泛应用。本研究建立了一种基于探针熔解曲线分析（MLMA）的多重连接反应（Multiple Ligation Reaction）方法，用于副溶血性弧菌57个K血清群的同时鉴定。根据418株包括39个K血清群的已知基因组序列和18个K血清群的公共数据库序列，获得了57个K血清群特异性基因序列，用于设计引物和探针。开发的用于鉴定副溶血性弧菌57 K血清群的MLMA检测显示出高重现性，检测内和检测间标准偏差和变异系数分别不超过1°C和1%。所有靶基因的检测限范围为0.1 - 1.0 ng/µl。我们通过双盲试验验证了MLMA检测试剂盒，可识别595 V。使用常规血清分型方法进行比较。结果表明，MLMA方法与传统血清学方法的Kappa值为0.936，与传统血清学方法的符合率为96.97%。此外，使用MLMA检测鉴定了5种罕见的K血清群，以及使用传统血清分型方法无法鉴定的18种菌株。因此，MLMA检测为副溶血性弧菌K血清群的分子血清分型提供了一种快速、稳健和有前景的工具，并具有潜在的应用于副溶血性弧菌感染暴发的检测和监测。
The serotyping of Vibrio parahaemolyticus, which is crucial to the surveillance and detection of outbreaks of vibriosis infection, has been widely used in many countries. In this study, we developed a molecular assay, named multiplex ligation reaction based on probe melting curve analysis (MLMA), for simultaneous identification of V. parahaemolyticus 57 K-serogroups. Based on the previous genomes of 418 strains including 39 K-serogroups and the 18 K-serogroups sequences from public databases, we obtained 57 K-serogroups specific gene sequences for designing primers and probes. The developed MLMA assay for identifying the V. parahaemolyticus 57 K-serogroups showed high reproducibility, with the intra- and inter-assay standard deviations and coefficients of variation of no more than 1°C and 1%, respectively. The limit of detection for all gene targets ranged from 0.1 to 1.0 ng/µl. We validated the MLMA assay with a double-blind test identifying 595 V. parahaemolyticus isolates using conventional serotyping methods for comparison. The results showed the kappa value between the MLMA assay and the traditional serological method was 0.936 and that there was a 96.97% consistency rate with conventional serotyping methods for all detected isolates. Additionally, five rare K-serogroups were identified using the MLMA assay, as well as 18 strains that could not be identified using the traditional serotyping method. Thus, the MLMA assay provides a rapid, robust, and promising tool for the molecular serotyping of V. parahaemolyticus K-serogroups and has the potential application to the detection of outbreaks and surveillance of V. parahaemolyticus infection.
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弧菌偏溶血：关于发病率，患病率和提高分子鉴定技术的综述。
影响因子：
5.2
作者：
Letchumanan V;Chan KG;Lee LH
通讯作者：
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影响因子：
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作者：
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DOI：
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发表时间：
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影响因子：
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影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
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