DEVELOPMENT OF REVERSE-TRANSCRIPTION LOOP-MEDIATED ISOTHERMAL AMPLIFICATION FOR AVIAN LEUKOSIS VIRUS SUBGROUPS A AND J

摘要

Neoplasic diseases in chickens are frequently caused by avian leukosis virus (ALV), including subgroup A (ALV-A) and subgroup J (ALV-J) mostly. The objective of the present study was to develop a one-step, single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) test for detecting ALV-A and ALV-J separately. Primers were designed from the conserved region of the pol and env genes of ALV-A and ALV-J for RT-LAMP tests. Other avian viruses were used to determine the specificityof these tests. The results showed that a ladder pattern on gel electrophoresis was observed for ALV-A and ALV-J specifically and no cross-reaction was observed with other avian viruses. Serial dilutions of ALV-A and ALV-J were used to evaluate the detection limits of these tests. The present RT-LAMP tests showed 10-100 times more sensitivity than the conventional reverse transcription polymerase chain reaction. Thus, these tests are rapid and accurate for the detection of ALV-A and ALV-J.