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目的对1例感染源不明的三日疟病例进行实验室检测分析并确诊。方法收集该病例的临床资料,并进行流行病学调查。对患者及其供血者的血样进行血涂片病原学检查、疟疾快速诊断检测(RDT)和巢式PCR检测,并对阳性结果进行测序比对。结果该患者无疟疾流行区居留史和既往疟疾感染史,有外科手术时大量输血史。外周血涂片镜检查见三日疟原虫。流行病学调查显示,该患者接受了3位供血者的血液,但3位供血者血样经血涂片镜检、RDT和巢式PCR检测均未查见疟原虫,后经改进的巢式-多重PCR检测及结果测序比对后显示,与其中1名非洲籍留学生供血者的阳性扩增条带具100%同源性。结论该病例因输血而感染三日疟。实验室检测疟疾疑难病例需经多方法验证方可确诊,改进的巢式-多重PCR方法对低密度疟原虫感染有较好的检测效果。
Objective To analyze and diagnose a case of Plasmodium malariae of unknown origin. Methods The clinical data of this case were collected and the epidemiological investigation was carried out. Blood samples from patients and their blood donors were tested for blood smear etiology, malaria rapid diagnostic test (RDT) and nested PCR test, and the positive results were sequenced and aligned. Results The patient had no history of malaria endemic area and previous malaria infection, and had a large number of transfusions during surgery. Plasmodium malaria parasite was detected in peripheral blood smear microscopy. Epidemiological surveys showed that the patient received blood from three blood donors. However, three blood donors were blood smear microscopy, none of the RDT and nested PCR tests showed plasmodia, and an improved nested-multiple PCR test and sequencing result showed that there was 100% homology with the positive amplification band of one of the blood donors from African-born students. Conclusion The case was infected with blood malaria on the 3rd day after transfusion. Laboratory tests for difficult cases of malaria require multi-method validation before diagnosis. The improved nested-multiplex PCR method has a good detection effect on low-density Plasmodium infection.