[1]吴 意,金 娴,樊春卉,等.单管双重巢式聚合酶链反应及基因测序技术鉴别呼吸道合胞病毒A,B亚型[J].现代检验医学杂志,2018,33(01):44-48,51.[doi:10.3969/j.issn.1671-7414.2018.01.001]
　WU Yi,JIN Xian,FAN Chun-hui,et al.Identification of Respiratory Syncytial Virus A and B SubtypesbyTube Double Nested Polymerase Chain Reaction and Gene Sequencing Technology[J].Journal of Modern Laboratory Medicine,2018,33(01):44-48,51.[doi:10.3969/j.issn.1671-7414.2018.01.001]
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单管双重巢式聚合酶链反应及基因测序技术鉴别呼吸道合胞病毒A,B亚型()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第33卷
期数:
2018年01期
页码:
44-48,51
栏目:
论著
出版日期:
2018-02-05

文章信息/Info

Title:
Identification of Respiratory Syncytial Virus A and B SubtypesbyTube Double Nested Polymerase Chain Reaction and Gene Sequencing Technology
文章编号:
1671-7414(2018)01-044-06
作者:
吴 意1; 金 娴1; 樊春卉1; 陆学东2; 赵 毅1
1.深圳市第七人民医院检验科,广东深圳 518081; 2.中山大学附属第八医院检验医学部,广东深圳 51803
Author(s):
WU Yi1; JIN Xian1; FAN Chun-hui1; LU Xue-dong2; ZHAO Yi1
1.Department of Clinical Laboratoory,Shenzhen Seven Peopel'sHospital, Guangdong Shenzhen 518081,China; 2.Department of Clinical Laboratoory,the Eighth Affiliated Hospital of Sun Yat-sen University,Guangdong Shenzhen 518033,China
关键词:
巢式PCR;  基因测序;  呼吸道合胞病毒;  亚型
分类号:
R373.14; Q503
DOI:
10.3969/j.issn.1671-7414.2018.01.001
文献标志码:
A
摘要:
目的 建立呼吸道合胞病毒(RSV)A,B亚型的鉴别方法,以供临床科研应用。方法 运用荧光定量聚合酶链反应技术(FQ-PCR)筛选出2015～2017年50例住院儿童咽拭子RSV阳性标本; 根据RSV的G蛋白编码基因核苷酸序列设计单管双重巢式PCR引物进行基因分型; 其A,B亚型产物经测序与GeneBank库中核苷酸序列进行比对分析鉴定; 采用卡方检验对结果进行统计学分析。结果 50例咽拭子RSV阳性患儿呼吸道合胞病毒A,B亚型及混合感染率分别为82.00%,14.00%和4.00%,差异有统计学显著性意义(χ2=81.06,P<0.01); RSV分型结果与基因测序结果完全符合。结论 深圳东部地区以A亚型流行为主,混合感染并存; 单管双重巢式聚合酶链反应(PCR)及基因测序技术适用于RSV的A,B亚型鉴别,具有灵敏度高、特异度强和准确率高的特点。
Abstract:
Abstract:Objective Objective to establish a method for identification of respiratory syncytial virus(RSV)A and B subtypes for clinicalresearch and application.Methods Using fluorescent quantitative polymerase chain reaction(FQ-PCR)screened 50 cases of throat swab that RSV were positive in hospitalized children from 2015 to 2017.The genotypingwas performed according to the nucleotide sequence of G protein coding gene,and a single tube double nested PCR primers was designed for it.A and B subgroupby sequencing to conduct comparative analysis with nucleotide sequence in the Genebank.The results were analyzed by chi-square test.Results In the 50 cases of throat swab RSV positive children,respiratory syncytial virus A and B subtype and mixed infection rates were 82.00%,14.00% and 4.00%,respectively.The difference was statistically significant(χ2=81.06,P<0.01).The RSV fractal results were consistent with the gene sequencing results.Conclusion The eastern part of shenzhen was dominated by respiratory syncytial virus A subtype epidemic and mixed infection.Single tube double nested polymerase chain reaction(PCR)and gene sequencing technique is suitable for theidentification of A and B subtypes of RSV.It is characterized by high sensitivity,specificity and high accuracy.

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备注/Memo

备注/Memo:
基金项目:深圳市盐田区科技局科技计划资助项目(20160102)。 作者简介:吴 意(1982-),男,大学本科,主管检验师,主要从事病原微生物分子诊断学研究,E-mail:845007571@qq.com。
更新日期/Last Update: 2018-02-07