[1]张薇,李丽娟,荣长仙,等.基于环介导恒温扩增和微流控芯片的生殖道支原体检测方法构建[J].中国皮肤性病学杂志,2021,(03):343-348.[doi:10.13735/j.cjdv.1001-7089.202004075 ]
 ZHANG Wei,LI Lijuan,RONG Changxian,et al.Construction of A Microfluidic Chip for Four Genital Mycoplasmas Based on Loop-mediated Isothermal Amplification[J].The Chinese Journal of Dermatovenereology,2021,(03):343-348.[doi:10.13735/j.cjdv.1001-7089.202004075 ]
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基于环介导恒温扩增和微流控芯片的生殖道支原体检测方法构建()
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《中国皮肤性病学杂志》[ISSN:1001-7089/CN:61-1197/R]

卷:
期数:
2021年03期
页码:
343-348
栏目:
检验技术
出版日期:
2021-03-01

文章信息/Info

Title:
Construction of A Microfluidic Chip for Four Genital Mycoplasmas Based on Loop-mediated Isothermal Amplification
文章编号:
1001-7089(2021)03-0343-06
作者:
张薇1李丽娟1荣长仙2方虹舒1
1.重庆市急救医疗中心检验科, 重庆 400014; 2.重庆市急救医疗中心妇产科, 重庆 400014
Author(s):
ZHANG Wei1 LI Lijuan1 RONG Changxian2 FANG Hongshu1
(1.Clinical Laboratory, Chongqing Emergency Medical Center, Chongqing 400014, China; 2.Department of Obstetrics and Gynecology, Chongqing Emergency Medical Center, Chongqing 400014, China)
关键词:
微流控芯片 环介导恒温扩增 解脲脲原体 微小脲原体 生殖支原体 人型支原体
Keywords:
Microfluidic chip Loop-mediated isothermal amplification Ureaplasma urealy-ticum Ureaplasma parvum Mycoplasma genitalium Mycoplasma hominis
分类号:
R 446.5
DOI:
10.13735/j.cjdv.1001-7089.202004075
文献标志码:
B
摘要:
目的 建立一种结合环介导恒温扩增和微流控芯片技术检测生殖道支原体的方法。方法 针对4种生殖道支原体包括解脲脲原体(UU)、微小脲原体(UP)、生殖支原体(MG)、人型支原体(MH)的DNA序列进行特异性引物设计和合成,制备微流控芯片,配制恒温扩增反应体系后进行芯片加样,于核酸分析仪中进行核酸扩增和结果判读,并对该芯片进行准确性、灵敏度、特异性和重复性验证。结果 成功制备的4种生殖道支原体的微流控芯片具有较好的准确性、灵敏度、特异性和重复性,整个检测过程在1 h内即可完成,UU、MG、MH的最低检测限为1 000拷贝/μL,而UP的最低检测限为5 000拷贝/μL。结论 基于环介导恒温扩增技术的4种生殖道支原体微流控芯片具有快速、准确、高通量等优点,在临床上具有广泛的应用前景。
Abstract:
Objective To establish a microfluidic chip method for four genital mycoplasmas based on loop-mediated isothermal amplification(LAMP).Methods Specific primers were designed and synthesized of four genital mycoplasmas including Ureaplasma urealyticum(UU), Ureaplasma parvum(UP), Mycoplasma genitalium(MG)and Mycoplasma hominis(MH).The microfluidic chip were constructed, the samples were amplificated by PCR.The accuracy, sensitivity, specificity and repeatability of the results were also verified.Results The microfluidic chip established successfully, and it had good accuracy, sensitivity, specificity and repeatability, and the detection process was finished within 1 hour.The detection limit of Ureaplasma urealyticum, Mycoplasma genitalium, Mycoplasma hominis were 1 000 copies/μL, while that of Ureaplasma parvum was 5 000 copies/μL.Conclusion The fast, accurate, high-throughput microfluidic chip method may has an extensive application for clinical diagnosis.

参考文献/References:

[1] Roachford OSE, Nelson KE, Mohapatra BR.Comparative genomics of four Mycoplasma species of the human urogenital tract: Analysis of their core genomes and virulence genes[J].Int J Med Microbiol, 2017, 307(8): 508-520.
[2] Wang QY, Li RH, Zheng LQ, et al.Prevalence and antimicrobial susceptibility of Ureaplasma urealyticum and Mycoplasma hominis in female outpatients, 2009-2013[J].J Microbiol Immunol Infect, 2016, 49(3): 359-362.
[3] 蒲清泉, 吴文耀, 杜丽, 等.液体培养法检测泌尿生殖道支原体感染的准确性评价[J].检验医学与临床, 2017, 14(18): 2694-2697.
[4] Cox C, McKenna JP, Watt AP, et al.Ureaplasma parvum and Mycoplasma genitalium are found to be significantly associated with microscopy-confirmed urethritis in a routine genitourinary medicine setting[J].Int J STD AIDS, 2016, 27(10): 861-867.
[5] 胡宏波,冯金,李晓明.男性生殖道解脲脲原体检测方法比较[J].广西中医学院学报,2009, 12(1): 27-29.
[6] 高峰, 华利忠, 于伯华, 等.鉴别牛5种重要病毒的LAMP-基因芯片方法的建立[J].动物医学进展, 2019, 40(5): 7-12.
[7] Ma YD, Luo K, Chang WH, et al.Amicrofluidic chip capable of generating and trapping emulsion droplets for digital loop-mediated isothermal amplification analysis[J].Lab Chip, 2018, 18(2): 296-303.
[8] 李艳燕, 高美丽, 赛亚飞, 等.呼吸道病原体核酸恒温扩增芯片十三联检在下呼吸道感染诊断中的价值[J/OL].中华肺部疾病杂志(电子版), 2019, 12(2): 171-174.
[9] Taylor-Robinson D.Mollicutes in vaginal microbiology: myco-plasma hominis, ureaplasma urealyticum, ureaplasma parvum and mycoplasma genitalium[J].Res Microbiol, 2017, 168(9): 875-881.
[10] Combaz-Sohnchen N, Kuhn A.A systematic review of mycoplasma and ureaplasma in urogynaecology[J].Geburtshilfe Frauenheilkd, 2017, 77(12): 1299-1303.
[11] 张岱, 刘朝晖.生殖道支原体感染诊治专家共识[J].中国性科学, 2016, 25(3): 80-82.
[12] Leon D, Retamal J, Silva R, et al.Molecular detection of sexually transmitted agents in a symptomatic group of men and its relationship with sexual behavior[J].Rev Chilena Infectol, 2016, 33(5): 505-512.
[13] Houssin T, Cramer J, Grojsman R, et al.Ultrafast, sensitive and large-volume on-chip real-time PCR for the molecular diagnosis of bacterial and viral infections[J].Lab Chip, 2016, 16(8): 1401-1411.
[14] 戈海泽,梁慧敏,卢晋英,等.基因芯片技术在常见肠道致病菌感染检测中的应用[J].山东医药, 2013, 53(4): 24-26.
[15] Li C, Ding XR, Liu ZZ, et al.Rapid identification of Candida spp.frequently involved in invasive mycoses by using flow-through hybridization and Gene Chip(FHGC)technology[J].J Microbiol Methods, 2017, 132: 160-165.

备注/Memo

备注/Memo:
[基金项目] 重庆市科学技术委员会资助项目(cstc2018jscx-msybX0107)
[通信作者] 方虹舒, E-mail: fangfangfhs@163.com
更新日期/Last Update: 2021-02-10