rna and dna bacteriophages as molecular diagnosis controls in clinical virology a comprehensive study of more than 45,000 routine pcr testsrna和dna噬菌体控制分子诊断在临床病毒学的全面研究超过45000常规pcr测试.pdfVIP

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rna and dna bacteriophages as molecular diagnosis controls in clinical virology a comprehensive study of more than 45,000 routine pcr testsrna和dna噬菌体控制分子诊断在临床病毒学的全面研究超过45000常规pcr测试.pdf

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rna and dna bacteriophages as molecular diagnosis controls in clinical virology a comprehensive study of more than 45,000 routine pcr testsrna和dna噬菌体控制分子诊断在临床病毒学的全面研究超过45000常规pcr测试

RNA and DNA Bacteriophages as Molecular Diagnosis Controls in Clinical Virology: A Comprehensive Study of More than 45,000 Routine PCR Tests 1,2 1,2 2 1 1 Laetitia Ninove *, Antoine Nougairede , Celine Gazin , Laurence Thirion , Ilenia Delogu , Christine Zandotti1,2, Remi N. Charrel1,2, Xavier De Lamballerie1,2,3 ´ ´ ´ ´ ˆ 1 UMR190, Universite de la Mediterranee and Institut de Recherche pour le Developpement, Marseille, France, 2 Pole de Microbiologie and Maladies Infectieuses, ˆ Assistance Publique – Hopitaux de Marseille, Marseille, France, 3 EHESP School of Public Health, Marseille, France Abstract Real-time PCR techniques are now commonly used for the detection of viral genomes in various human specimens and require for validation both external and internal controls (ECs and ICs). In particular, ICs added to clinical samples are necessary to monitor the extraction, reverse transcription, and amplification steps in order to detect false-negative results resulting from PCR-inhibition or errors in the technical procedure. Here, we performed a large scale evaluation of the use of bacteriophages as ICs in routine molecular diagnosis. This allowed to propose simple standardized procedures (i) to design specific ECs for both DNA and RNA viruses and (ii) to use T4 (DNA) or MS2 (RNA) phages as ICs in routine diagnosis. Various technical formats for using phages as ICs were optimised and validated. Subsequently, T4 and MS2 ICs were evaluated in routine real-time PCR or RT-PCR virological diagnostic tests, using a series of 8,950 clinical samples (representing 36 dis