基于18S rRNA基因的芜湖市疟疾病例溯源研究

Malaria tracing in Wuhu City based on the 18S rRNA gene

目的研究芜湖市疟疾病例疟原虫种系发育和地理聚类。方法采集2018年8月至2019年9月皖南医学院附属弋矶山医院收治的4例疟疾患者血样,提取血样疟原虫DNA,采用巢式PCR技术对其18S rRNA基因进行扩增、克隆测序分析,并与GenBank中疟原虫相应序列进行比较分析,采用MEGA X软件建树,运用分子进化树分析芜湖市疟疾的地理起源。结果 4例患者PCR电泳结果获得片段均为206 bp,经BLAST比对证实均为恶性疟原虫感染。系统进化树显示,4例病例检测基因片段与GenBank中非洲分离株聚集在相同亚支,其余由GenBank上获取的恶性疟原虫序列也都分别按照地理来源聚集在相同的进化亚支内。结论 18S rRNA基因是能够确定疟疾感染来源地的准确、特异性的溯源分子标记物。

To study the development and geographical clustering of malaria cases in Wuhu city,blood samples were collected from four patients with imported malaria admitted to Yijishan Hospital affiliated to Wannan Medical College from August 2018 to September 2019,and genomic DNA was extracted.The 18S rRNA genes of the malaria parasite were amplified by nested PCR technology,and the gene sequence of the amplified product was obtained by cloning and sequencing.MEGA X software was used to construct a phylogenetic tree to analyze the geographical origin of the malaria cases.The electrophoresis results of the PCR products from the four patients were 206 bp and confirmed by BLAST comparison.The phylogenetic tree showed that the four detected gene fragments were clustered in the same subbranch as GenBank African isolates,and the other Plasmodium falciparum sequences obtained from GenBank were clustered in the same evolutionary subbranch according to geographical sources.This study demonstrated that the 18S rRNA gene is an accurate and specific traceability molecular marker that can determine the origin of infection in patients with malaria.