An adhesion molecule-based colloidal gold immunochromatography assay strip for rapidly and specifically detecting chicken antibodies against Mycoplasma gallisepticum.
The fragment of the VlhA1.2 gene was cloned from a Mycoplasma gallisepticum (MG) DNA library by serial PCRs after the same-sense mutagenesis of three TGA codons encoding tryptophan (Trp). Following transforming the generated plasmid of pKG-VlhA1.2, the recombinant VlhA1.2-GST fusion protein of 92kDa was induced and recognized by anti-MG sera. After GST-affinity chromatographic purification, the VlhA-based colloidal gold immunochromatography assay (GICA) strips were generated. The GICA strips specifically detected anti-MG antibodies, but not antibodies against Mycoplasma synoviae and other positive sera against non-MG pathogens tested. The GICA strips were 128-fold more sensitive to detect anti-MG antibodies, as compared with traditional serological methods and were stored at 4° C for 15 months without loss of their sensitivity and specificity. Analysis of sample revealed that, the GICA strips were highly sensitive, specific and stable for the on-site surveillance of MG infections by unskilled users.
Zhang
Jinliang
J
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
Guo
Yan
Y
Hu
Sishun
S
Bi
Dingren
D
Wang
Xiliang
X
Xiao
Yuncai
Y
Gao
Xiaocong
X
Li
Zili
Z
Liu
Mei
M
Peng
Fuhu
F
Xu
Qingrong
Q
D000372:Agglutination Tests / Q000662:veterinary; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D015153:Blotting, Western / Q000662:veterinary; D002645:Chickens / Q000276:immunology / Q000382:microbiology*; D002846:Chromatography, Affinity / Q000379:methods / Q000662:veterinary*; D017979:Gold Colloid; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D045686:Mycoplasma gallisepticum / Q000276:immunology*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011201:Poultry Diseases / Q000175:diagnosis* / Q000276:immunology; D011933:Reagent Kits, Diagnostic / Q000382:microbiology / Q000662:veterinary; D011934:Reagent Strips; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D017979:Gold Colloid; D011933:Reagent Kits, Diagnostic; D011934:Reagent Strips; D011994:Recombinant Proteins