Possible use of RNA isolate from inactivated vaccine for external positive control in reverse transcription-based detection of foot-and-mouth disease virus in bull semen.
This study has demonstrated the novel use of inactivated and purified vaccine against FMD virus for detection and analysis. RNA isolate was efficiently generated from the vaccine for an external positive control for reverse transcription polymerase chain reaction (RT-PCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays. The target DNA fragment sequences from the 2B region and 3D RNA polymerase gene of the virus for RT-PCR and RT-LAMP respectively were successfully amplified using the RNA template. Laboratories lacking complex equipment may not be feasible to handle high-risk viruses for conventional methods such as the isolation and culture of live viruses. Here, with the use of these molecular tools, novel use of RNA isolate from inactivated, purified vaccine proved to be an effective external positive control for the assays. Therefore, with these methods, the derived RNA control template aids in a safe method for screening FMD virus for diagnostic laboratories. And by using the same technique, it is then possible to generate a standard for diagnosing any other infectious viral diseases.
Paraguison
Rubigilda C
RC
Molecular Genetics Laboratory, Genetic Improvement Program Unit, Philippine Carabao Center, Science City of Muñoz, Nueva Ecija 3119, Philippines. rubigee@gmail.com
Flores
Ester B
EB
Cruz
Libertado C
LC
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D002417:Cattle; D005536:Foot-and-Mouth Disease / Q000175:diagnosis*; D029544:Foot-and-Mouth Disease Virus / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*; D012661:Semen / Q000821:virology*; D015164:Vaccines, Inactivated / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014765:Viral Vaccines / Q000235:genetics*
D000956:Antigens, Viral; D012367:RNA, Viral; D015164:Vaccines, Inactivated; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C103557:virus-infection-associated antigen, Foot-and-mouth disease virus