机构：[1]Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, National Center for Children’s Health, Capital Medical University, Beijing, China临床科室科研平台职能科室耳鼻咽喉头颈外科临床流行病与循证医学中心儿科研究所首都医科大学附属北京儿童医院[2]Qingdao Center for Disease Control and Prevention, Qingdao, China[3]Liaoning Provincial Center for Disease Control and Prevention, Shenyang, China
Background In this study, we aimed to optimize the condition of propidium monoazide (PMA) treatment for direct detection of Mycobacterium tuberculosis (MTB) from clinical specimens. Methods The light exposure time, dark incubation time, bacterial load, and PMA concentration were varied to determine the optimal condition of PMA treatment. Results Overall, the maximum Delta Cq value was observed in the group receiving a light exposure time of 20 minutes, which was significantly higher than the others (P < 0.05). The prolongation of dark incubation time seemed more likely to result in greater Delta Cq value, and the Delta Cq values were 2.0, 4.1, 6.5, 10.1, and 12.7 cycles under dark incubation time of 10, 20, 40, 60, and 120 minutes, respectively. Alternatively, the 4+ samples exhibited favorable detection results at the application of 10(4)-fold dilution by PMA assay with Cq values higher than 35 cycles. Further evaluation revealed that the PMA assay showed an accordance rate of 98.0% (98/100) among clinical sputa. Conclusions we develop an acceptable method to directly identify the live bacteria from sputum samples. Our data demonstrate that the dark incubation plays a crucial role in the efficacy of PMA treatment for MTB.
基金：
Capitals Funds for Health Improvement and Research [2016-4-1141]; Beijing Health System Top Level Health Technical Personnel Training Plan [20153079]
第一作者机构：[1]Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, National Center for Children’s Health, Capital Medical University, Beijing, China[*1]Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing, China.
通讯作者：
通讯机构：[1]Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, National Center for Children’s Health, Capital Medical University, Beijing, China[3]Liaoning Provincial Center for Disease Control and Prevention, Shenyang, China[*1]Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing, China.[*2]Liaoning Provincial Center for Disease Control and Prevention, Shenyang, China.
推荐引用方式(GB/T 7714)：
Lu Jie,Zheng Huiwen,Chu Ping,et al.Direct detection from clinical sputum samples to differentiate live and dead Mycobacterium Tuberculosis[J].JOURNAL OF CLINICAL LABORATORY ANALYSIS.2019,33(3):-.doi:10.1002/jcla.22716.
APA：
Lu, Jie,Zheng, Huiwen,Chu, Ping,Han, Shujing,Yang, Hui...&Yang, Zuosen.(2019).Direct detection from clinical sputum samples to differentiate live and dead Mycobacterium Tuberculosis.JOURNAL OF CLINICAL LABORATORY ANALYSIS,33,(3)
MLA：
Lu, Jie,et al."Direct detection from clinical sputum samples to differentiate live and dead Mycobacterium Tuberculosis".JOURNAL OF CLINICAL LABORATORY ANALYSIS 33..3(2019):-
