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用于疟疾检测和疟原虫种类鉴定的巢式聚合酶链反应

[Nested PCR for malaria detection and Plasmodium species identification].

作者信息

Shi Yong-xia, Huang Ji-cheng, Su Jin-kun, Hong Ye, Li Xiao-bo, Zheng Kui, Xing Lu-qin, Guo Bo-xuan

机构信息

Health Quarantine Laboratory, Guangdong Inspection and Quarantine Technology Center, Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou 510700, China.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2011 Aug;29(4):263-6.

Abstract

According to the sequences of small subunit ribosomal RNA (SSU rRNA) gene of Plasmodium spp., universal and species-specific primers were designed to detect malaria and identify species. 60 blood samples were detected by the established nested PCR method. The results were compared with those of microscopic examination. 40 blood samples were Plasmodium-positive by nested PCR with 22 samples of P. falciparum, 13 of P. vivas, 3 with P. falciparum and P. vivax mixed infection, 1 of P. ovale and 1 of unclassified malaria infection. Altogether, the coincidence between the results of nested PCR and microscopy stood for 76.7% (46/60), including 18 of P. falciparum, 11 of P. vivax and 17 negatives. Further sequence analysis and real-time PCR were performed to detect blood samples with discrepancy, results of which were the same as that of nested PCR. The amplified product of P. ovale was sequenced and showed 100% homology to the corresponding part of P. ovale SSU rRNA gene sequence (GenBank No. DQ845247), which confirmed that the case was imported ovale malaria.

摘要

根据疟原虫小亚基核糖体RNA(SSU rRNA)基因序列,设计通用引物和种特异性引物用于检测疟疾并鉴定虫种。采用建立的巢式PCR方法对60份血样进行检测,并将结果与显微镜检查结果进行比较。巢式PCR检测出40份血样疟原虫阳性,其中恶性疟原虫22份,间日疟原虫13份,恶性疟原虫和间日疟原虫混合感染3份,卵形疟原虫1份,未分类疟疾感染1份。巢式PCR结果与显微镜检查结果的总符合率为76.7%(46/60),其中恶性疟原虫符合18份,间日疟原虫符合11份,阴性符合17份。对结果有差异的血样进一步进行序列分析和实时PCR检测,结果与巢式PCR相同。对卵形疟原虫的扩增产物进行测序,结果显示与卵形疟原虫SSU rRNA基因序列(GenBank登录号:DQ845247)的相应部分有100%的同源性,证实该病例为输入性卵形疟。

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