一种高效准确定量计数噬菌体肽库目标分子的方法  

An Efficient and Accurate Method for Counting Target Molecules in Phage-Display Peptide Library

作　　者：王可耕[1] 曾庆仁[2] 禹正杨[3] 曾铁兵[4] 刘彦[1,2] 

机构地区：[1]南华大学医学院寄生虫学教研室,衡阳421001 [2]中南大学医学细胞与分子生物学实验中心,长沙410013 [3]南华大学附属第一医院肿瘤外科,衡阳421001 [4]南华大学医学院微生物学和免疫学教研室,衡阳421001

出　　处：《中国寄生虫学与寄生虫病杂志》2011年第3期236-238,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.30570952);湖南省卫生厅项目(2009);湖南省科技项目(2010)~~

摘　　要：利用双层琼脂平板法与荧光实时定量PCR(Real-time PCR)技术分别测定少量、中量和大量噬菌体的效价。结果显示,双层琼脂平板法与Real-time PCR法均可准确测定噬菌体效价。双层琼脂平板法所得符合条件的平板约为总数的1/3,需重复测定10次,每次消耗噬菌体10μl,测得噬菌体效价的组内变异系数为4.93%～30.38%。Real-timePCR法重复测定3次即可,每次消耗噬菌体1μl,其组内变异系数仅为0.02%～0.25%,明显低于双层琼脂平板法。提示,Real-time PCR技术可简单、迅速地测定噬菌体效价。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 μl 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 μl 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.

关 键 词：噬菌体 效价 双层琼脂平板法 荧光实时定量PCR 

分 类 号：R392.33[医药卫生—免疫学]