[1]刘爱玲,刘淑燕,刘爱胜,等.人偏肺病毒基因芯片检测方法的建立及初步应用[J].现代检验医学杂志,2017,32(06):19-22.[doi:10.3969/j.issn.1671-7414.2017.06.001]
　LIU Ai-ling,LIU Shu-yan,LIU Ai-sheng,et al.Establishment and Preliminary Application of the Gene Chip Detection Method of Human Pneumosis Virus[J].Journal of Modern Laboratory Medicine,2017,32(06):19-22.[doi:10.3969/j.issn.1671-7414.2017.06.001]
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人偏肺病毒基因芯片检测方法的建立及初步应用()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第32卷
期数:
2017年06期
页码:
19-22
栏目:
论著
出版日期:
2017-12-25

文章信息/Info

Title:
Establishment and Preliminary Application of the Gene Chip Detection Method of Human Pneumosis Virus
文章编号:
1671-7414(2017)06-019-04
作者:
刘爱玲1; 刘淑燕1; 刘爱胜1; 刘 军2; 朱彩云1; 郭龙华1
1.深圳市龙华区人民医院检验科, 广东深圳 518109; 2.广东医科大学病原生物学实验室,广东湛江 524023
Author(s):
LIU Ai-ling1; LIU Shu-yan1; LIU Ai-sheng1; LIUJun2; ZHU Cai-yun1; GUO Long-hua1
1.Department of Clinical Laboratory,Shenzhen Longhua District People's Hospital,Guangdong Shenzhen 518109,China; 2.Pathogen Biology Laboratory,Guangdong Medical University,Guangdong Zhanjiang 524023,China
关键词:
人偏肺病毒;  基因芯片;  常规反转录酶-聚合酶链锁反应;  建立;  应用
分类号:
Q503; R373.1
DOI:
10.3969/j.issn.1671-7414.2017.06.001
文献标志码:
A
摘要:
目的 建立深圳地区人偏肺病毒(human metapneumovirus,hMPV)核苷酸的快速、特异和敏感的基因芯片检测方法,为hMPV检测和诊断提供有效的诊疗手段。方法 针对hMPV病毒高度保守区域基因序列,应用引物设计软件Primer5设计hMPV的荧光PCR引物; 应用探针设计软件Array Designer4.20设计hMPV的寡核苷酸检测探针,将寡核苷酸探针点样至醛基玻片上制备hMPV基因芯片,并与常规反转录酶-聚合酶链锁反应(RT-PCR)平行比较,对其灵敏度、特异度和重复性,以及用于临床样本的适用性等进行评价。结果 hMPV基因芯片法检测hMPV的特异度为96.23%(230/239),灵敏度可达2.0×101个/μl,线性范围为2.0×101～2.0×107个/μl,且重复性好; 初步检测深圳地区300份临床鼻咽拭子标本,基因芯片法检出率为20.3%(61/300),明显高于常规RT-PCR法的9.7%(29/300),两方法检测的灵敏度之间差异有统计学意义(χ2=39.205,P<0.05); 两种方法检测结果一致性一般(kappa=0.360 7)。结论 建立了hMPV基因芯片检测法,基因芯片法检测hMPV具有很高的特异度和灵敏度,且检测线性范围广,为实验室开展hMPV的流行监测和早期诊断提供了新的检测技术。
Abstract:
Abstract:Objective To establish human metapneumovirus(hMPV)nucleotide rapid,specific and sensitive gene chip detection method,and provide effective diagnostic methods for hMPV detection and diagnosis in shenzhenarea.Methods The fluorescence PCR primer of hMPV was designed for the highly conservative regional gene sequence of hMPV virus.Application of array probe design software designer 4.20 hMPV oligonucleotide detectionprobe design,oligonucleotide probe sample points to aldehyde slides on the preparation of hMPV gene chip,and parallel compared with conventional reverse transcriptase-polymerase chain reaction(RT-PCR),the sensitivity,specificity andrepeatability,and was used to evaluate the clinical applicability of the samples.Results hMPV gene chip method to detect hMPV specificity was 96.23%(230/239),the sensitivity was 2.0×101/μl,linear range was2.0×101～2.0×107/μl,and the repeatability was good.Initial tests inshenzhen area 300 clinical specimens on a nasopharyngeal swab,gene chip methoddetection rate was 20.3%(61/300),significantly higher than the conventional RT-PCR method 9.7%(29/300),the difference was statistically significant between the sensitivity of two methods(χ2=39.205,P<0.05).The results wereconsistent in two methods(kappa=0.360 7).Conclusion Established hMPV microarray assay,gene chip method to detect hMPV has high specificity and sensitivity,wide linear range and detection.The popularity of hMPVmonitoring for laboratory provides a new detecting technology and early diagnosis.

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备注/Memo

备注/Memo:
基金项目:广东医科大学校院联合科研基金立项项目(编号:L2016008)。 作者简介:刘爱玲(1983-),女,医学硕士,主管技师,主要从事微生物和分子生物学检验工作,E-mail:liuailing1983@163.com。
更新日期/Last Update: 2017-12-26