Jafari Sayeh, Le Bras Jacques, Bouchaud Olivier, Durand Rémy
Centre National de Référence pour la Chimiosensibilité du Paludisme, Université Paris V et Assistance Publique-Hôpitaux de Paris, Hôpital Bichat-Claude Bernard, Laboratoire de Parasitologie, Paris, France.
J Infect Dis. 2004 Jan 15;189(2):195-203. doi: 10.1086/380910. Epub 2004 Jan 8.
We have developed a new fragment-analysis method to enumerate the clones and to quantify their proportions within Plasmodium falciparum isolates. We prospectively enrolled 20 adult patients with uncomplicated malaria who were returning to France from various sub-Saharan countries, from January 2000 through July 2001. The analysis of clonal populations was performed on blood samples obtained at 10 times: 1 before treatment with oral quinine and 9 during the first 96 h of the treatment. The resistance genotypes pfcrt and dhfr were determined for chloroquine and antifolinics. Multiple P. falciparum genotypes were detected in 19 (95%) of 20 patients: 2, 3, 4, and 5 genotypes were found in 4, 9, 4 and 2 patients, respectively. Disappearance and reappearance of some clones within a few hours was observed. Individual clones represented 0.4%-99.4% of total parasitemia. Surprisingly, in 10 of 15 subjects tested, resistance genotypes varied according to the time of blood collection. These findings may have important implications with regard to the interpretations of resistance studies.
我们开发了一种新的片段分析方法，用于在恶性疟原虫分离株中计数克隆并量化其比例。2000年1月至2001年7月，我们前瞻性地招募了20名从撒哈拉以南非洲各国返回法国的患单纯性疟疾的成年患者。对在10个时间点采集的血样进行克隆群体分析：口服奎宁治疗前1次，治疗的前96小时内9次。确定了氯喹和抗叶酸药物的抗性基因型pfcrt和dhfr。20名患者中有19名（95%）检测到多种恶性疟原虫基因型：分别在4、9、4和2名患者中发现了2、3、4和5种基因型。观察到一些克隆在数小时内消失和重新出现。单个克隆占总寄生虫血症的0.4%-99.4%。令人惊讶的是，在15名受试对象中的10名中，抗性基因型随采血时间而变化。这些发现可能对抗性研究的解释具有重要意义。
