Development and validation of an immunogold chromatographic test for on-farm detection of PRRSV.
An immunochromatographic test strip was developed to detect porcine reproductive and respiratory syndrome virus (PRRSV). The test uses two gold-labeled monoclonal antibodies: D5 against recombinant nucleocapsid protein (rN) and E9 against recombinant M protein (rM). In the test, PRRSV binds to a mixture of D5 and E9 labeled with colloidal gold; the complexes move through a membrane and are captured by rabbit anti-rM and anti-rN antibodies at a test line, producing a reddish-purple band because of the increased concentration of gold. Unbound monoclonal antibodies move past the test line to be captured by goat anti-mouse antibodies, producing a band at a control line. In samples without PRRSV or with low virus concentration, a band appears only at the control line. A crossover-trial demonstrated that the test strip was highly specific for PRRSV. The test strip detection limit was between 7.8x10(3) and 1.6x10(4) TCID(50)/ml. Analysis of 100 clinical samples indicated that the sensitivity, specificity, and accuracy of the immunochromatographic test strip relative to reverse transcription polymerase chain reaction (RT-PCR) were 97.0, 93.9, and 96.0%, respectively. Because the test is simple and rapid, it can be used by an unskilled person to detect PRRSV in the field.
Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, China.
Sheng-hua
SH
Zhou
Shang-jin
SJ
Cui
Chang-mu
CM
Chen
Fan-chao
FC
Zhang
Jun
J
Li
Shun
S
Zhou
Jin-Sik
JS
Oh
D000818:Animals; D000911:Antibodies, Monoclonal; D002845:Chromatography / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis*; D019316:Porcine respiratory and reproductive syndrome virus / Q000302:isolation & purification*; D011817:Rabbits; D012680:Sensitivity and Specificity; D013552:Swine
D000911:Antibodies, Monoclonal