Development of a simple and rapid immunochromatographic strip test for diarrhea-causative porcine rotavirus in swine stool.
A rapid and simple immunochromatography (IC) strip test, for specific detection of porcine rotavirus (PRV) in stool specimen, was developed. Monoclonal antibodies (mAbs) to the OSU strain of PRV have been produced in mice. Among them, two hybridoma clones that generate mAb-1 and mAb-2, respectively, specific for VP6 protein of PRV, have been selected. In the IC configuration, mAb-1, one of the selected mAbs was used to the designed coat microparticles (MP), while another mAb-2 was used to fix it on the nitrocellulose membrane strip to form a result line. The control line was formed on the same membrane strip past the result line by fixing anti-mouse IgG antibody. The IC test was capable of detecting 1000 plaque-forming units of PRV/ml in less than 5min, and the binding capacity was demonstrated by specific recognition of PRV only, but not other porcine diarrhea viruses, transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV). The IC test produced positive results with all the nine PRV-positive stool specimens and negative results with five different non-PRV specimens, which were identified previously by the polymerase chain reaction (PCR) test, respectively. The results indicate an excellent concordance between the two methods, suggesting a potential application of the three combinated IC tests (PRV, TGEV and PEDV) for the on-site, rapid screening of porcine diarrhea cases.
Kang
J H
JH
Hanrib Lifetech, Inc., Venture Bldg., Bio21 Foundation Center, 305-6 Jinju-City, KyeongNam, Republic of Korea.
Kwon
D H
DH
Chung
T W
TW
Kim
Y D
YD
Lee
H G
HG
Kim
J W
JW
Choe
I S
IS
Kim
K-W
KW
Lim
J S
JS
Song
E Y
EY
Kim
C-H
CH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D003967:Diarrhea / Q000662:veterinary* / Q000821:virology; D005243:Feces / Q000821:virology*; D007118:Immunoassay / Q000379:methods*; D011934:Reagent Strips; D012401:Rotavirus / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000821:virology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011934:Reagent Strips