高通量基因芯片技术在登革病毒检测和分型中的应用
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摘要
目的建立能同时检测登革病毒4个亚型的基因芯片方法,并进行验证。方法使用建立的高通量快速基因芯片检测28份疑似登革热病例血清样本,与商品化荧光RT-PCR试剂盒检测结果进行比较,分析方法的结果一致性;将4个已知型别的登革病毒阳性样本混合,使用建立的芯片平台进行检测,评价该芯片在登革病毒分型中的应用价值。结果 28份疑似登革热病例的样本经高通量基因芯片检测,18份为登革病毒阳性,与商品化实时荧光RT-PCR试剂盒结果一致,符合率100%;混合样本中登革病毒4个型别均可检出。结论建立的芯片检测技术平台在登革热诊断方面具有较高的应用价值,值得在疾病预防、卫生检疫领域推广。
Objective To evaluate an integrated lab-on-chip method constructed for rapid identification and typing of dengue virus. Methods The high-throughput lab-on-chip method was established by designing primers and probes of dengue virus. 28 samples of suspected dengue fever cases were used to analyze the consistency of the lab-on-chip method and commercial Real-time RT-PCR kit. The typing application for dengue virus of the lab-onchip method was evaluated by using the mixed samples of four dengue virus subtypes. Results There were 18 among 28 suspected dengue fever specimens were positive by the lab-on-chip method, which was consistent with the commercial Real-time RT-PCR kit, and the accordance rate was 100%. The four subtypes of dengue virus in the mix samples could be differentiated exactly. Conclusion This integrated lab-on-chip method had higher application value in the diagnosis of dengue fever and is worth popularizing in the field of disease prevention and health quar-antine.
Objective To evaluate an integrated lab-on-chip method constructed for rapid identification and typing of dengue virus. Methods The high-throughput lab-on-chip method was established by designing primers and probes of dengue virus. 28 samples of suspected dengue fever cases were used to analyze the consistency of the lab-on-chip method and commercial Real-time RT-PCR kit. The typing application for dengue virus of the lab-onchip method was evaluated by using the mixed samples of four dengue virus subtypes. Results There were 18 among 28 suspected dengue fever specimens were positive by the lab-on-chip method, which was consistent with the commercial Real-time RT-PCR kit, and the accordance rate was 100%. The four subtypes of dengue virus in the mix samples could be differentiated exactly. Conclusion This integrated lab-on-chip method had higher application value in the diagnosis of dengue fever and is worth popularizing in the field of disease prevention and health quar-antine.
引文
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[15] Conceicao TM,Da Poian AT,Sorgine MH.A real-time PCR procedure for detection of dengue virus serotypes 1,2,and 3,and their quantitation in clinical and laboratory samples[J]. J Virol Methods,2010,163(1):1-9.
[2] Messina JP,Brady OJ,Scott TW,et al. Global spread of dengue virus types:mapping the 70 year history[J]. Trends Microbiol,2014,22(3):138-146.
[3] Guzman A,Isturiz RE. Update on the global spread of dengue[J]. Int J Antimicrob Agents,2010,36(S1):40-42.
[4] Grange L,Simon LE,Sakuntabhai A,et al. Epidemiological risk factors associated with high global frequency of inapparent dengue virus infections[J]. Front Immunol,2014(5):280.
[5]吴忠华,罗鹏,徐琦,等.登革病毒检测技术研究进展[J].生物技术通讯,2013(5):727-731.
[6] Wang SM,Sekaran SD. Early diagnosis of dengue infection using a commercial dengue duo rapid test kit for the detection of NS1,IGM,and IGG[J]. Am J Trop Med Hyg,2010,83(3):690-695.
[7] Sayers EW,Barrett T,Benson DA,et al. Database resources of the National Center for Biotechnology Information[J]. Nucleic Acids Res,2012. 40(Database issue):13-25.
[8]杜建伟,潘先海.中国登革热流行概况与流行特征[J].中华流行病学杂志,2010,31(12):1429-1433.
[9] Zhang H,Li Z,Lai S,et al. Evaluation of the performance of a dengue outbreak detection tool for China[J]. PLoS One,2014,9(8):e106144.
[10] Teles FS. Biosensors and rapid diagnostic tests on the frontier between analytical and clinical chemistry for biomolecular diagnosis of dengue disease:a review[J]. Anal Chim Acta,2011,687(1):28-42.
[11] Zaytseva NV,Montagna RA,Baeumner AJ. Microfluidic biosensor for the serotype-specific detection of dengue virus RNA[J].Anal Chem,2005,77(23):7520-7527.
[12] Ngo HT,Wang HN,Fales AM,et al. DNA bioassay-on-chip using SERS detection for dengue diagnosis[J]. Analyst,2014,139(22):5655-5665.
[13] Tan JJ,Capozzoli M,Sato M,et al. An integrated lab-on-chip for rapid identification and simultaneous differentiation of tropical pathogens[J]. PLoS Negl Trop Dis,2014,8(7):e3043.
[14] Neeraja M,Lakshmi V,Lavanya V,et al.Rapid detection and differentiation of dengue virus serotypes by NS1 specific reverse transcription loop-mediated isothermal amplification(RTLAMP)assay in patients presenting to a tertiary care hospital in Hyderabad,India[J]. J Virol Methods,2015(211):22-31.
[15] Conceicao TM,Da Poian AT,Sorgine MH.A real-time PCR procedure for detection of dengue virus serotypes 1,2,and 3,and their quantitation in clinical and laboratory samples[J]. J Virol Methods,2010,163(1):1-9.