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Zastosowanie Rt-PCR W Czasie Rzeczywistym
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ISBN10: 6203897310
ISBN13: 9786203897319
Publisher: Wydawnictwo Nasza Wiedza
Published: Apr 24 2025
Pages: 68
Weight: 0.23
Height: 0.16 Width: 6.00 Depth: 9.00
Language: Polish
ISBN13: 9786203897319
Publisher: Wydawnictwo Nasza Wiedza
Published: Apr 24 2025
Pages: 68
Weight: 0.23
Height: 0.16 Width: 6.00 Depth: 9.00
Language: Polish
Wirus pryszczycy (FMDV) jest diagnozowany za pomocą testu izolacji wirusa w polączeniu z testem ELISA antygenu, a ostatnio jednoetapowym RT-PCR. Zastosowanie fluorogenicznego RT-PCR zyskuje coraz większe znaczenie w szybkim wykrywaniu FMDV z próbek klinicznych, które nie mogą byc przetwarzane za pomocą zwyklych testów izolacji wirusa i antygenu ELISA. Ilościowy RT-PCR w czasie rzeczywistym (qRT-PCR) lączy odwrotną transkrypcję z amplifikacją PCR i wykrywaniem pozytywnej amplifikacji przy użyciu sond fluorescencyjnych, takich jak sonda TaqMan (R). W niniejszym badaniu, qRT-PCR oparty na sondzie TaqMan(R) zostal ustandaryzowany i wykorzystany do wykrywania genomu FMDV z próbek klinicznych, w tym wydzielin nosowych, osocza i plynów ustno-gardlowych (próbki probang). Wstępną standaryzację przeprowadzono przy użyciu różnych seryjnych rozcieńczeń izolatu FMDV OTNN 24/84 (106,8 TCID50), a 102 wirusy można bylo zidentyfikowac za pomocą TaqMan(R) qRT-PCR. Standardy RNA zostaly zsyntetyzowane in vitro z plazmidu zawierającego 110 par zasad regionu 3D izolatu FMDV typu O. Dziesięciokrotne seryjne rozcieńczenie RNA zostalo wykonane i użyte jako standardowy RNA do wygenerowania krzywych wzorcowych.
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