Detection of foot and mouth disease and porcine reproductive and respiratory syndrome viral genes using microarray chip.
Two viral pathogens, namely, porcine reproductive and respiratory syndrome virus (PRRSV) and foot and mouth disease virus (FMDV), were selected as models for multiple pathogen detection in a cDNA microarray. Two signature regions selected from ORF2 (around 500 bp) and ORF5 (around 600 bp) of PRRVS (America serotype), and one signature region from structural genes VP1 (around 500 bp) of FMDV type O were designed and spotted on a nylon membrane. For PCR sensitivity study, the cloned FMDV-VP1 template could be diluted to near one copy and its PCR product was still detectable in gel electrophoresis. In the microarray detection, the labelling FMDV probes (3 mg/ml) could be diluted 320 times and still maintained a visible colour when hybridized with the chip. Using the mixing primers, the microarray chip demonstrated rapid and accurate detection of the specific genes. To our knowledge, this preliminary study is the first example reported applying the long signature sequences to the multiple pathogen detection in cDNA microarray.
Liu
Y-C
YC
Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan. ycliu@dragon.nchu.edu.tw
Huang
G S
GS
Wu
M-C
MC
Hong
M-Y
MY
Hsiung
K-P
KP
D000818:Animals; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics* / Q000302:isolation & purification; D007091:Image Processing, Computer-Assisted; D020411:Oligonucleotide Array Sequence Analysis / Q000662:veterinary*; D016366:Open Reading Frames / Q000235:genetics; D010865:Pilot Projects; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology*; D015678:Viral Structural Proteins / Q000235:genetics
D012367:RNA, Viral; D015678:Viral Structural Proteins