Real-time quantitative PCR for determining the burden of Plasmodium falciparum parasites during pregnancy and infancy
Real-time quantitative PCR for determining the burden of Plasmodium falciparum parasites during pregnancy and infancy
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DOI：
10.1128/jcm.43.8.3630-3635.2005
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发表时间：
2005-08-01
影响因子：
9.4
通讯作者：
King, CL
中科院分区：
文献类型：
--
作者：
Malhotra, I;Dent, A;King, CL
实时定量聚合酶链式反应(RTQ-PCR)提供了一种快速、准确和重复性好的寄生虫定量方法。然而，实时定量-聚合酶链式反应对预测疟疾临床结果的价值尚不清楚。在这里，我们将实时定量-聚合酶链式反应(RTQ-PCR)与血涂片镜检、套式聚合酶链式反应(NPCR)和寄生虫循环抗原(CAG)检测进行比较，以检测肯尼亚孕妇及其婴儿的恶性疟原虫，并将这些结果与其新生儿的产次和出生体重相关(n=554)。在检测孕期疟疾方面，nPCR是最敏感的方法，其次是RTQ-PCR、CAG和血涂片。初产妇母亲外周血中疟疾感染率(46%)高于经产妇(35%；P&0.001)，寄生虫负担差异12倍(几何平均值分别为25,870和2,143个扩增/亩L血；P<；0.0001)。同样，通过RTQ-PCR确定的胎盘疟疾的存在在初产妇女中大约是经产妇女的两倍(21%比13%；P<；0.01)。RTQ-PCR估计孕妇中疟疾感染的存在和强度与低出生体重婴儿密切相关，特别是在扩增子数量较高的婴儿中。实时定量-聚合酶链式反应确定了血液涂片漏掉的感染疟疾的妇女，她们的后代有体重不足的风险。相比之下，nPCR和CAG检测发现的疟疾与产次或低出生体重的相关性要弱得多。因此，实时定量-聚合酶链式反应提供了比血液涂片更敏感的寄生虫负担估计，并可预测孕妇和新生儿感染疟疾的临床结果。
Real-time quantitative PCR (RTQ-PCR) provides a quick, accurate, and reproducible quantification of parasites. However, the value of RTQ-PCR for predicting clinical outcomes of malaria is unknown. Here, we compared RTQ-PCR to microscopy of blood smears, nested PCR (nPCR), and parasite circulating-antigen (CAg) assays for detection of Plasmodium falciparum in pregnant Kenyan women and their infants and related these findings to parity and birth weights in their newborns (n = 554). nPCR was the most sensitive assay for detection of malaria in pregnancy, followed in decreasing order of sensitivity by RTQ-PCR, CAg assays, and blood smears. RTQ-PCR detected a higher frequency of malaria infection (46%) in maternal peripheral blood in primiparous than in multiparous women (35%; P < 0.001), with a > 12-fold difference in parasite burden (geometric mean = 25,870 versus 2,143 amplicons/mu l blood; P < 0.0001). Similarly, the presence of placental malaria determined by RTQ-PCR was approximately twofold higher in primiparous versus multiparous women (21% versus 13%; P < 0.01). The presence and intensity of malaria infection in pregnant women estimated by RTQ-PCR strongly correlated with low-birth-weight babies, especially in those with high amplicon numbers. RTQ-PCR identified malaria-infected women, missed by blood smear, who were at risk for having underweight offspring. By contrast, malaria detected by nPCR and CAg assay showed a much weaker association with parity or low birth weight. Thus, RTQ-PCR provides an estimate of parasite burden that is more sensitive than blood smear and is predictive of clinical outcomes of malaria infection in pregnant women and newborns.