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实时定量 PCR 研究抗利什曼原虫药物在印度内脏利什曼病中的寄生虫动力学。

Study of parasite kinetics with antileishmanial drugs using real-time quantitative PCR in Indian visceral leishmaniasis.

机构信息

Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi-221005, UP, India.

出版信息

J Antimicrob Chemother. 2011 Aug;66(8):1751-5. doi: 10.1093/jac/dkr185. Epub 2011 May 24.

Abstract

OBJECTIVES

This study describes parasite kinetics in the blood of visceral leishmaniasis patients treated with liposomal amphotericin B (L-AmB) or a preformed fat emulsion of amphotericin B (ApL) using real-time quantitative PCR (qPCR).

METHODS

Forty-six patients were treated with a single dose (15 mg/kg of body weight) of either L-AmB (n = 13) or ApL (n = 33). qPCR was used to estimate parasite kinetics by detection of Leishmania donovani DNA using kinetoplast DNA-specific primers in peripheral blood samples using an absolute quantification method.

RESULTS

The mean parasite load decreased from baseline (day 0) values of 894.07 and 980.48 to 71.72 and 211.52 parasite genomes/mL at day 7 in L-AmB and ApL groups, respectively, and at day 30 these further declined to 8.30 and 133.98 parasite genomes/mL, respectively. At day 30 post-treatment evaluation, the decline in parasite load was significantly greater (P = 0.024) with L-AmB compared with ApL. Four of 33 patients in the ApL group failed treatment (1 primary failure and 3 relapses) with the presence of parasites, whereas all patients in the L-AmB group were cured at 6 month follow-up.

CONCLUSIONS

qPCR can be a tool to measure parasite dynamics accurately and provide a marker to measure the efficacy of various drugs. It can be used as a test of cure, allowing us to do away with invasive and risky methods such as splenic or bone marrow aspiration.

摘要

目的

本研究使用实时定量 PCR(qPCR)描述了用脂质体两性霉素 B(L-AmB)或两性霉素 B 预形成脂肪乳剂(ApL)治疗内脏利什曼病患者血液中的寄生虫动力学。

方法

46 名患者接受了单剂量(15mg/kg 体重)的 L-AmB(n=13)或 ApL(n=33)治疗。qPCR 用于通过使用绝对定量方法在外周血样本中使用动基体 DNA 特异性引物检测利什曼原虫 DNA 来估计寄生虫动力学。

结果

寄生虫负荷平均值从基线(第 0 天)值 894.07 和 980.48 分别下降到第 7 天的 L-AmB 和 ApL 组的 71.72 和 211.52 寄生虫基因组/mL,第 30 天进一步下降到 8.30 和 133.98 寄生虫基因组/mL。在治疗后 30 天评估时,L-AmB 组与 ApL 组相比,寄生虫负荷下降显著更大(P=0.024)。ApL 组 33 名患者中有 4 名治疗失败(1 例原发性失败和 3 例复发)且存在寄生虫,而 L-AmB 组所有患者在 6 个月随访时均治愈。

结论

qPCR 可以成为一种准确测量寄生虫动态的工具,并提供一种测量各种药物疗效的标志物。它可以用作治愈的测试,使我们能够避免进行脾脏或骨髓抽吸等侵入性和危险的方法。

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