A quantitative reverse transcription-polymerase chain reaction for detection of Getah virus. 2018

Sing-Sin Sam, and Boon-Teong Teoh, and Cheah-Mun Chee, and Noor-Adila Mohamed-Romai-Noor, and Juraina Abd-Jamil, and Shih-Keng Loong, and Chee-Sieng Khor, and Kim-Kee Tan, and Sazaly AbuBakar
Tropical Infectious Diseases Research and Education Centre (TIDREC), University of Malaya, Kuala Lumpur, Malaysia.

Getah virus (GETV), a mosquito-borne alphavirus, is an emerging animal pathogen causing outbreaks among racehorses and pigs. Early detection of the GETV infection is essential for timely implementation of disease prevention and control interventions. Thus, a rapid and accurate nucleic acid detection method for GETV is highly needed. Here, two TaqMan minor groove binding (MGB) probe-based quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays were developed. The qRT-PCR primers and TaqMan MGB probe were designed based on the conserved region of nsP1 and nsP2 genes of 23 GETV genome sequences retrieved from GenBank. Only the qRT-PCR assay using nsP2-specific primers and probe detected all two Malaysia GETV strains (MM2021 and B254) without cross-reacting with other closely related arboviruses. The qRT-PCR assay detected as few as 10 copies of GETV RNA, but its detection limit at the 95% probability level was 63.25 GETV genome copies (probit analysis, P ≤ 0.05). Further validation of the qRT-PCR assay using 16 spiked simulated clinical specimens showed 100% for both sensitivity and specificity. In conclusion, the qRT-PCR assay developed in this study is useful for rapid, sensitive and specific detection and quantification of GETV.

UI MeSH Term Description Entries
D006734 Horse Diseases Diseases of domestic and wild horses of the species Equus caballus. Equine Diseases,Disease, Equine,Disease, Horse,Diseases, Equine,Diseases, Horse,Equine Disease,Horse Disease
D006736 Horses Large, hoofed mammals of the family EQUIDAE. Horses are active day and night with most of the day spent seeking and consuming food. Feeding peaks occur in the early morning and late afternoon, and there are several daily periods of rest. Equus caballus,Equus przewalskii,Horse, Domestic,Domestic Horse,Domestic Horses,Horse,Horses, Domestic
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000524 Alphavirus A genus of TOGAVIRIDAE, also known as Group A arboviruses. They are serologically related to each other and are transmitted by mosquitoes. The type species is the SINDBIS VIRUS. Arboviruses, Group A,Barmah forest virus,Getah virus,Sagiyama virus,Alpha Virus,Alphaviruses,Alpha Viruses,Arbovirus, Group A,Group A Arbovirus,Group A Arboviruses
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D012463 Saliva The clear, viscous fluid secreted by the SALIVARY GLANDS and mucous glands of the mouth. It contains MUCINS, water, organic salts, and ptylin. Salivas
D013552 Swine Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA). Phacochoerus,Pigs,Suidae,Warthogs,Wart Hogs,Hog, Wart,Hogs, Wart,Wart Hog
D013553 Swine Diseases Diseases of domestic swine and of the wild boar of the genus Sus. Disease, Swine,Diseases, Swine,Swine Disease
D057230 Limit of Detection Concentration or quantity that is derived from the smallest measure that can be detected with reasonable certainty for a given analytical procedure. Limits of Detection,Detection Limit,Detection Limits

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