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目的应用多重巢式PCR技术对一例输入性疟疾混合感染病例进行诊断和鉴定。方法采集镜检初诊为输入性卵形疟患者血样,进行疟疾快速诊断试剂检测、显微镜镜检复核、多重巢式PCR检测及测序分析。结果该患者经快速诊断试剂检测结果为非恶性疟原虫阳性;镜检复核查见寄生于红细胞内的疟原虫环状体、大滋养体和配子体,根据形态鉴定为卵形疟;多重巢式PCR产物经琼脂糖凝胶电泳,在760bp和205bp处有特异性扩增条带,测序后经Blast比对,分别与Gen Bank数据库中卵形疟原虫变异亚种和恶性疟原虫部分序列的一致性为99.00%。结论该患者经多重巢式PCR和序列分析确诊为湖北省首例输入性卵形疟原虫变异亚种与恶性疟原虫混合感染病例。
Objective To diagnose and identify a case of mixed malaria infection with multiplex nested PCR. Methods The first visit of microscopic examination was blood samples of imported ovipositive malaria patients. The samples were tested for rapid diagnosis of malaria, microscopic examination, multiple nested PCR and sequencing analysis. Results The patient was tested positive for non-Plasmodium falciparum by rapid diagnostic reagent. Plasmodium larvae, trophozoites and gametocytes parasitized in erythrocytes were found by microscopic examination and were identified as oval-shaped malaria according to morphology. Multiple nested PCR The products were amplified by agarose gel electrophoresis with specific bands at 760bp and 205bp. After sequencing, they were aligned with Blast, and were consistent with the partial sequences of Plasmodium falciparum variant and Plasmodium falciparum in Gen Bank database Is 99.00%. Conclusions This patient was confirmed by multiple nested PCR and sequence analysis as the first case of mixed infection of Plasmodium ovale and Plasmodium falciparum in Hubei Province.