目的 研究二代测序技术（NGS）在美洲大蠊携带细菌检测中的应用，建立一种可检测美洲大蠊体内、体表所携带未知种类细菌的方法，分析不同环境下美洲大蠊携带细菌情况及其与环境的关系。方法 2016年8－9月，以中山地区居民区、港口和医院环境中捕获的美洲大蠊为材料，对其16S rDNA V1～V9可变区分析，选取V3～V4可变区作为目的基因，利用NGS对其携带的细菌种类进行检测和分析。结果 港口、居民区和医院采集的美洲大蠊分别携带1 564、1 786和1 779个运算分类单位；利用NGS分别可鉴定出细菌123属36种、131属42种和136属48种，细菌种类数超过传统方法。通过对样品进行物种多样性和聚类分析发现，同一环境下的样本无法完全聚集在一起。结论 16S rDNA V3～V4可变区适合对美洲大蠊体内、体表携带的细菌进行检测，利用NGS可一次性检出多种细菌；其检出的细菌种类远超出传统方法且操作简便；美洲大蠊携带细菌的种类与其生存环境无明显的区域特异性。
Objective Establish an efficient method for detection of bacteria carried by Periplaneta americana both inside and outside with next generation sequencing (NGS) platform. The P. americana were collected from different environment, and the relationship between the P. americana-carrying bacteria to its living environment was analyzed as well. Methods During August and September in 2016, P. americana were collected from the port, hospital and residential areas of Zhongshan, 16S rDNA V3-V4 was selected as the target gene. Bacteria carried by P. americana both inside and outside was studied by NGS method. Results Periplaneta americana collected at the port area were identified 1 564 OTU, 123 genera, 36 species of bacteria; 1 779 OTU, 136 genus, 48 species of bacteria from the P. americana collected at the hospital area, 1 786 OTU, 131 genus, 42 species of bacteria from the P. americana collected at the residential area. The number of bacteria detected by NGS method was more than that of traditional methods. Through the analysis of species diversity and cluster analysis, the samples in the same environment cannot be aggregated completely. Conclusion 16S rDNA V3-V4 variable region was suitable for detection of bacteria carried by P. americana, NGS technology can be used to detect a variety of pathogenic bacteria of P. americana both inside and outside. Compared with traditional methods, the bacterial species detected by NGS were far beyond the traditional methods and were easy to operate; By cluster analysis of the samples, it has no obvious relationship between the bacteria carried by P. americana to its living environment.
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