[1]敖知,郭述良.噬菌体Chy2生物扩增法的建立及其检测效果的评价[J].第三军医大学学报,2013,35(03):241-244.
 Ao Zhi,Guo Shuliang.Establishment of phage amplified biologically assay using phage Chy2 and evaluation of its capacity of detecting Mycobacterium Tuberculosis in sputum samples[J].J Third Mil Med Univ,2013,35(03):241-244.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
35卷
期数:
2013年第03期
页码:
241-244
栏目:
论著
出版日期:
2013-02-15

文章信息/Info

Title:
Establishment of phage amplified biologically assay using phage Chy2 and evaluation of its capacity of detecting Mycobacterium Tuberculosis in sputum samples
作者:
敖知郭述良
重庆医科大学附属第一医院呼吸与危重症医学科
Author(s):
Ao Zhi Guo Shuliang
Department of Respiratory and Critical Care Medicine, First Affiliated Hospital, Chongqing Medical University, Chongqing, 400016, China
关键词:
分枝杆菌噬菌体Chy2结核分枝杆菌灵敏度特异性
Keywords:
mycobacteriophage Chy2 Mycobacterium tuberculosis sensitivity specificity
分类号:
R373.9; R378.911; R446.5
文献标志码:
A
摘要:
目的      建立噬菌体Chy2生物扩增法,并评价其检测临床痰标本结核分枝杆菌的应用价值。       方法        噬菌斑测试法观察噬菌体杀灭剂灭活Chy2的工作浓度和作用时间;噬菌斑计数法判断Chy2的最佳感染时间和感染滴度;分别用噬菌体Chy2和D29生物扩增法检测58份临床痰标本,以改良罗氏培养法作诊断标准,对比评价Chy2和D29的检测准确性。       结果        6%硫酸亚铁铵溶液作用5 min可彻底灭活游离Chy2;Chy2滴度为2.2×1011CFU/mL时,与结核分枝杆菌孵育4 h的感染效率最高;噬菌体Chy2生物扩增法对痰标本的检测灵敏度、特异性、Youden指数、阳性预测值和阴性预测值分别为94.7%、85.0%、0.797、0.923、0.895,噬菌体D29生物扩增法的检测结果相应为76.3%、95.0%、0.713、0.967、0.678,噬菌体Chy2生物扩增法的灵敏度、阴性预测值优于噬菌体D29生物扩增法(P<0.05),两者特异性、Youden指数、阳性预测值无统计学差异(P>0.05)。      结论        成功建立了噬菌体Chy2生物扩增法,Chy2较D29检测特异性相当,灵敏度有所提高。
Abstract:
Objective        To explore the methodology of the phage amplified biologically assay (PhaB assay) using a newly isolated and identified mycobacteriophage named Chy2, and to evaluate its clinical value for detecting Mycobacterium tuberculosis (MTB) in sputum samples.       Methods        The optimal concentration and time of phage-inactivation agents were observed by spot tests. The optimal time and titer of phage to infect MTB were confirmed by plaque counts. Fifty-eight clinical sputum samples were collected from patients suspected with tuberculosis (TB) with no previous history of TB treatment, and were detected by PhaB assay using phage Chy2 and D29, respectively. Modified Lowenstein-Jensen (L-J) culture method was used as the diagnostic standard.        Results        Ferrous ammonium sulfate (FAS) solution (6%) mixed with Chy2 for 5 min was able to completely inactivate the extracellular phages. Chy2 of 2.2×1011CFU/mL could infect MTB most effectively after incubating with MTB for 4 hours. PhaB assay using Chy2 to detect MTB in sputum samples only needed 48 hours, and had sensitivity, specificity, Youden index, positive predictive value (PPV) and negative predictive value (NPV) of 94.7%, 85.0%, 0.797, 0.923 and 0.895, respectively, while the counterparts of D29 PhaB assay were 76.3%, 95.0%, 0.713, 0.967 and 0.678, respectively. Chy2-PhaB assay had a higher sensitivity and NPV than D29-PhaB assay did (P<0.05).       Conclusion        PhaB assay using Chy2 has been successfully established with a similar specificity and a higher sensitivity for detecting MTB in sputum as compared with D29, and has the potential to be a valuable, rapid and economic screening approach for detecting pulmonary TB.

参考文献/References:

敖知, 郭述良. 噬菌体Chy2生物扩增法的建立及其检测效果的评价[J].第三军医大学学报,2013,35(3):241-244.

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更新日期/Last Update: 2013-01-29