Detection of Newcastle disease virus using a SYBR Green I real time polymerase chain reaction.
A two-step SYBR Green I real time polymerase chain reaction (PCR, real time PCR) for the detection of Newcastle disease virus (NDV) was developed. A melting curve analysis was performed to distinguish specific from non-specific products and primer dimers. Regardless of different virus pathotypes the melting temperature (Tm) ranged from 86 degrees C to 87 degrees C. The sensitivity of the real time PCR was compared with the reverse transcription (RT)-nested PCR enzyme-linked immunosorbent assay (ELISA, RT-nested PCR ELISA). Whereas the detection limit of the real time PCR was 10 pg DNA, the RT-nested PCR ELISA and conventional PCR could only detect up to 1 ng and 10 ng DNA, respectively. Thus the real time PCR offers a sensitive, rapid and convenient method for screening large number of NDV specimens.
Tan
S W
SW
Faculty of Veterinary Medicine, University Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia.
Omar
A R
AR
Aini
I
I
Yusoff
K
K
Tan
W S
WS
D000818:Animals; D052160:Benzothiazoles; D002645:Chickens / Q000821:virology; D003959:Diamines; D005456:Fluorescent Dyes; D009521:Newcastle Disease / Q000175:diagnosis* / Q000821:virology; D009522:Newcastle disease virus / Q000235:genetics / Q000302:isolation & purification*; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction; D011804:Quinolines; D012680:Sensitivity and Specificity
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I