A real-time polymerase chain reaction-based test system for quantitation of Gumboro disease virus | Тест-система на основе ПЦР в реальном времени для количественного определения вируса болезни Гамборо
2010
Pogoda, A.A. | Potekhina, M.A.
A real-time polymerase chain reaction-based test system for quantification of infectious bursal disease (Gumboro disease) virus was developed. The reaction parameters were analyzed, which affected the linear relationship of a C1 depentanizer to the quantity of cDNA. The use of specific primers for reverse transcription was shown to have some advantage over that of random hexanucleotides. As was found, with low concentrations of RNA the number of cDNA copies when using hexanucleotides reduced disproportionately to the number of RNA, and to obtain an adequate result, it was necessary to reduce the concentrations of primers. At the same time, using specific primers provided the required proportionality. Using high concentrations of a mixture of RNA comprising not only viral RNA but also cellular RNA sometimes resulted in reducing the efficiency of reverse transcription. It is concluded that the mentioned efficicnce of producing cDNA by using specific primers allows standardizing the analysis techniques of material with no regard to the concentrations of virap particles in a sample and its specificity. The test-system as developed is recommended to conduct monitoring of infectious bursal disease both in field and natural conditions
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