[Validation of nested RT-PCR method for detection of measles virus in clinical samples].
The aim of this study was to perform the validation of nested RT-PCR method for detection of measles virus genome in laboratory of Virology Department (NIZP-PZH) conditions. The PCR reactions were made with specific primers for nucleoprotein gene. The validation of nested RT-PCR assay includes determination of limit of detection (LOD), specificity, intra- and inter-assay precision and influence of materials from clinical samples on the PCR reaction. The detection limit of nested RT-PCR was 255-510 RNA MeV/ml. The obtained results reveal high precision and lack of substances present in clinical samples influence on the PCR reaction. The application of nested RT-PCR assay for viral genomes other than MeV gave negative results. The obtained results indicated that the nested RT-PCR fulfilled validation criteria and can be used for routine laboratory diagnostics of measles virus.
Makówka
Agata
A
Zakład Wirusologii NIZP-PZH w Warszawie. amakowka@pzh.gov.pl
Siennicka
Joanna
J
D004279:DNA, Viral / Q000302:isolation & purification; D006801:Humans; D008459:Measles virus / Q000302:isolation & purification*; D010614:Pharynx / Q000382:microbiology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*; D014556:Urine / Q000382:microbiology
D004279:DNA, Viral