Loop-mediated isothermal amplification for rapid detection and differentiation of wild-type pseudorabies and gene-deleted virus vaccines.
A loop-mediated isothermal amplification (LAMP) assay was developed specifically for detection and differentiation of pseudorabies virus (PRV). One group of primers was designed to detect wild-type strains (i.e., strains with the gE gene) and the other group of primers was designed to detect both PRV gE-vaccine and wild-type strains (i.e., strains with the gG gene and with or without the gE gene). After amplification by Bst enzyme at a constant temperature of 65 degrees C, a laddering of bright products was visible following electrophoresis on a 2% agarose gel. LAMP was 100-1000-fold more sensitive than the standard PCR. The assay was specific in that it did not amplify other porcine viruses including porcine parvovirus, porcine circovirus type 1, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, classical swine fever virus, swine transmissible gastroenteritis coronavirus, and porcine epidemic diarrhea virus. Because of its sensitivity, specificity, and simplicity, the LAMP assay could be a useful method for early and rapid differentiation of swine vaccinated with PRV gE-deleted vaccine from swine infected with wild virus.
Zhang
Chao-Fan
CF
Division of Swine Infectious Disease, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of CAAS, Harbin 150001, China.
Cui
Shang-Jin
SJ
Zhu
Chao
C
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D017353:Gene Deletion; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011557:Pseudorabies / Q000175:diagnosis* / Q000821:virology; D022421:Pseudorabies Vaccines / Q000235:genetics; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D014613:Vaccines, Attenuated / Q000235:genetics
D017931:DNA Primers; D004279:DNA, Viral; D022421:Pseudorabies Vaccines; D014613:Vaccines, Attenuated