Detection of Plasmodium falciparum malaria parasites in vivo by real-time quantitative PCR
Detection of Plasmodium falciparum malaria parasites in vivo by real-time quantitative PCR
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DOI：
10.1016/s0166-6851(01)00379-6
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发表时间：
2001-12-01
影响因子：
1.5
通讯作者：
Sauerwein, RW
中科院分区：
文献类型：
--
作者：
Hermsen, CC;Telgt, DSC;Sauerwein, RW
基于18S核糖体RNA基因OFP的实时定量聚合酶链式反应(RTQ-PCR)。恶性疟原虫NF54被用来评估5名被蚊子实验感染的人类志愿者的寄生虫血症水平[荷兰；日期未给出]。所有5名志愿者在平均9.1天(范围为7.6-10.3天)后均出现寄生虫血症，通过厚涂片确定，并伴有轻度至中度症状的发热。经氯喹治疗后，感染的临床病程不复杂，症状在2-3天内消失，厚涂片显示原虫血症在2天内消失。用实时定量聚合酶链式反应，循环P。5名志愿者中有4名在感染后6.3天即检出恶性寄生虫(寄生虫密度：20-147只/毫升血)。在潜伏期内，疟疾寄生虫的数量稳步增加，在发现第一批寄生虫后20-24小时出现暂时但显著的下降，平均最大寄生虫数约为每毫升血液9,000只寄生虫(范围为1485-18 264)，在氯喹治疗开始后下降。厚涂片检测到的寄生虫数量与白细胞数量进行了校正，与RTQ-PCR估计的数量有很好的相关性(R=0.82)。结果表明，该方法是一种特异、灵敏的检测疟疾原虫的定量方法。
A novel real-time quantitative polymerase chain reaction (RTQ-PCR), based on the 18S ribosomal RNA genes ofP. falciparumNF54, was used to evaluate the level of parasitaemia in 5 human volunteers experimentally infected by mosquitoes [Netherlands; date not given]. All 5 volunteers developed parasitaemia, as determined by thick smear after a mean of 9.1 days (range: 7.6-10.3), and fever accompanied by mild to moderate symptoms. After anti-malaria treatment with chloroquine, the clinical course of the infection was uncomplicated, with resolution of the symptoms within 2-3 days and clearance of parasitaemia within 2 days as detected by thick smear. With the RTQ-PCR, circulatingP. falciparumparasites were detected in 4 of the 5 volunteers as early as 6.3 days after infection (parasite densities: 20-147 per ml blood). During the incubation period, the number of malaria parasites steadily increased, with a temporary but significant decrease at 20-24 h after detection of the first parasites, to a mean maximum of about 9000 parasites per ml of blood (range: 1485-18 264) and dropped after the start of chloroquine treatment. The number of parasites detected by thick smears was corrected for the number of leukocytes and showed a good correlation with the numbers estimated by the RTQ-PCR (R=0.82). These results demonstrate that this RTQ-PCR is a specific and sensitive quantitative assay for malaria parasite detection.