Muhamad Poonuch, Chacharoenkul Wanna, Rungsihirunrat Kanchana, Ruengweerayut Ronnatrai, Na-Bangchang Kesara
Pharmacology and Toxicology Unit, Graduate Program in Biomedical Sciences, Thammasat University, Thailand.
Asian Pac J Trop Biomed. 2011 Jan;1(1):49-53. doi: 10.1016/S2221-1691(11)60067-1.
To compare the applicability of the SYBR Green-I assay with the standard schizont maturation assay, for determination of sensitivity of Plasmodium vivax (P. vivax) to chloroquine and a new antifolate WR 99210.
The study was conducted at Mae Tao Clinic for migrant workers, Tak Province during April 2009 to July 2010. A total of 64 blood samples (1 mL blood collected into sodium heparinized plastic tube) were collected from patients with mono-infection with P. vivax malaria prior to treatment with standard regimen of a 3-day chloroquine. In vitro sensitivity of P. vivax isolates was evaluated by schizont maturation inhibition and SYBR Green-I assays.
A total of 30 out of 64 blood samples collected from patients with P. vivax malaria were successfully analyzed using both the microscopic schizont maturation inhibition and SYBR Green-I assays. The failure rates of the schizont maturation inhibition assay (50%) and the SYBR Green-I assay (54%) were similar (P=0.51). The median IC10s, IC50s and IC90s of both chloroquine and WR99210 were not significantly different from the clinical isolates of P. vivax tested. Based on the cut-off of 100 nM, the prevalences of chloroquine resistance determined by schizont maturation inhibition and SYBR Green-I assays were 19 and 11 isolates, respectively. The strength of agreement between the two methods was very poor for both chloroquine and WR99210.
On the basis of this condition and its superior sensitivity, the microscopic method appears better than the SYBR Green-I Green assay for assessing in vitro sensitivity of fresh P. vivax isolates to antimalarial drugs.
比较SYBR Green-I检测法与标准裂殖体成熟检测法在测定间日疟原虫对氯喹和新型抗叶酸药物WR 99210敏感性方面的适用性。
研究于2009年4月至2010年7月在北碧府为农民工开设的湄索诊所进行。在用3日氯喹标准疗法治疗前，从单纯感染间日疟原虫的患者中总共采集了64份血样（1毫升血液收集到含肝素钠的塑料试管中）。通过裂殖体成熟抑制试验和SYBR Green-I检测法评估间日疟原虫分离株的体外敏感性。
从间日疟原虫患者采集的64份血样中，共有30份成功通过显微镜下裂殖体成熟抑制试验和SYBR Green-I检测法进行了分析。裂殖体成熟抑制试验（50%）和SYBR Green-I检测法（54%）的失败率相似（P=0.51）。氯喹和WR99210的IC10、IC50和IC90中位数与所检测的间日疟原虫临床分离株无显著差异。基于100 nM的临界值，通过裂殖体成熟抑制试验和SYBR Green-I检测法确定的氯喹耐药率分别为19株和11株。两种方法在氯喹和WR99210方面的一致性都很差。
基于这种情况及其更高的敏感性，在评估新鲜间日疟原虫分离株对抗疟药物的体外敏感性方面，显微镜检测法似乎优于SYBR Green-I检测法。
