Preliminary application of Tem-PCR combined with luminex for detection of four common respiratory vi-ruses
10.16571/j.cnki.1008-8199.2016.09.015
- VernacularTitle:靶序列富集多重 PCR-液相芯片联合检测4种常见呼吸道病毒的初步应用
- Author:
Jie WANG
;
Weiping WANG
;
Yuan HU
;
Ning SUN
;
Bo YANG
;
Zhengkun XIA
;
Xiaojun LI
- Publication Type:Journal Article
- Keywords:
Respiratory viruses;
Target enriched multiplex PCR;
Luminex method
- From:
Journal of Medical Postgraduates
2016;29(9):958-963
- CountryChina
- Language:Chinese
-
Abstract:
Objective Respiratory viruses are the most common pathogens to cause respiratory tract infection in infants and children.The aim of the study was to establish a luminex-based molecular assay for rapid detection of four kinds of common respiratory viruses and provide measures for effective prevention and control . Methods 120 throat swab samples from patients with acute respiratory tract infection were collected in our hospital as disease group.30 normal specimens were used as control group .Specific up-stream and downstream primers , hybridization probes and super prim-ers were designed on the basis of conserved sequences of Influenza A and B viruses( FluA, FluB), respiratory syncytial virus types A and B ( RSVA, RSVB ) from available respiratory-virus sequence data-base.Recombinant plasmid and in vitro transcription RNA positive reference substances were established respectively .The testing sys-tem of Tem-PCR combined with luminex xMAP was built by amplification and optimization of hybridization .Comparative analysis were made between the detection results of the above method and those of single viral gene real -time PCR assay and luminex xTAG assay re-spectively. Results Rapid molecular assay was established to specifically detect the four kinds of respiratory viruses (FluA, FluB, RSVA and RSVB) with the sensitivity of 10 copies/μL.Rapid molecular assay and single viral real-time PCR assay were utilized to de-tect the throat swabs ( n=120 ) from suspected patients , the positive result of the former was 31 .7% ( 38/120 ) and the latter was 29.2%(35/120).The consistency test result indicated the two methods were consistent without a significant difference (k>0.7). Several samples were detected by luminex xTAG assay simultaneously , in which good consistency and significant difference were found in two assays by statistical analysis (k>0.6). Conclusion Preliminary clinical application has confirmed the novel molecular assay is sensitive, specific and rapid in simultaneous detection of FluA , FluB, RSVA and RSVB respiratory viruses , which provides experi-mental basis for accurate diagnosis of infected pathogens at early clinical stage .
