Wang Ke-Geng, Zeng Qing-Ren, Yu Zheng-Yang, Zeng Tie-Bing, Liu Yan
Department of Parasitology, School of Medicine, University of South China, Hengyang 421001, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2011 Jun;29(3):236-8.
The phage titer of samples representing the low, intermediate and high phage number was respectively determined by the double-layer agar plate (DLAP) method and real-time PCR assay. The two methods accurately measured the titer of samples. The plaques from about 1/3 double-agar layer plates could be used to determine the phage titer. The DLAP experiment should repeat 10 times with 10 microl sample each time, while the within-assay coefficient variation (CV) was 4.93%-30.38%. At the same time, the real-time PCR assay only repeated 3 times with 1 microl phage each time, while CV for within-assay ranged from 0.02% to 0.25%. Results indicated that real-time PCR is a simple and quick method for determining bacteriophage titer.
分别采用双层琼脂平板(DLAP)法和实时荧光定量PCR检测法测定代表低、中、高噬菌体数量的样本的噬菌体效价。这两种方法均能准确测定样本的效价。约1/3双层琼脂平板上的噬菌斑可用于测定噬菌体效价。DLAP实验每次用10微升样本应重复10次,实验内变异系数(CV)为4.93%-30.38%。同时,实时荧光定量PCR检测法每次仅用1微升噬菌体重复3次,实验内CV范围为0.02%-0.25%。结果表明,实时荧光定量PCR是一种测定噬菌体效价的简单快速方法。