Routine use of duplex real-time PCR assays including a commercial internal control for molecular diagnosis of opportunistic DNA virus infections

被引：17

作者：

Burrel, Sonia ^{[1
,2
]}

Fovet, Christelle ^{[1
]}

Brunet, Christel ^{[1
]}

Ovaguimian, Lydia ^{[1
]}

Hamm, Nathalie ^{[1
]}

Conan, Francoise ^{[1
]}

Kalkias, Laurence ^{[1
]}

Agut, Henri ^{[1
,2
]}

Boutolleau, David ^{[1
,2
]}

机构：

[1] Grp Hosp Pitie Salpetriere, AP HP, Serv Virol, F-75013 Paris, France

[2] UPMC Univ Paris 06, DETIV ER1, F-75013 Paris, France

来源：

JOURNAL OF VIROLOGICAL METHODS | 2012年 / 185卷 / 01期

关键词：

Laboratory-developed PCR; Internal control; Extraction and amplification process monitoring; Validity improvement; Quality assurance; HERPES-SIMPLEX; HUMAN-HERPESVIRUS-8; DNA; QUANTITATION;

D O I：

10.1016/j.jviromet.2012.05.031

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The aim of this work was to improve the validity of laboratory-developed real-time PCR protocols implemented in the laboratory for molecular diagnosis of opportunistic DNA virus infections using the Simplexa (TM) extraction and amplification control (SEAC) which allows the monitoring of the whole extraction and amplification process. Herpes simplex virus (HSV), varicella-zoster virus (VZV), human cytomegalovirus (CMV), Epstein-Barr virus (EBV), BK virus (BKV), and adenovirus (AdV) genomes were investigated in 152 different clinical specimens. The use of the SEAC did not influence the results of the different virus-specific PCRs. The SEAC results showed high reproducibility with a mean Cp value of 31.08 +/- 1.44, and were not influenced by the virus-specific PCR performed or the type of clinical specimen tested. The SEAC in the DNA extracts showed high stability during storage at both +4 degrees C and -20 degrees C. These data allowed establishing a new procedure for the validation of viral PCR results. In conclusion, the SEAC provides a reliable option for improving the diagnosis of opportunistic viral infections by laboratory-developed real-time PCR assays in quality assurance programs. (C) 2012 Elsevier B.V. All rights reserved.

引用

页码：136 / 141

页数：6

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