title: Development and evaluation of a one-step SYBR-Green I-based real-time RT-PCR assay for the detection and quantification of Chikungunya virus in human, monkey and mosquito samples.
abstract: This paper reports the development of a one-step SYBR-Green I-based realtime RT-PCR assay for the detection and quantification of Chikungunya virus (CHIKV) in human, monkey and mosquito samples by targeting the E1 structural gene. A preliminary evaluation of this assay has been successfully completed using 71 samples, consisting of a panel of negative control sera, sera from healthy individuals, sera from patients with acute disease from which CHIKV had been isolated, as well as monkey sera and adult mosquito samples obtained during the chikungunya fever outbreak in Malaysia in 2008. The assay was found to be 100-fold more sensitive than the conventional RT-PCR with a detection limit of 4.12x10(0) RNA copies/μl. The specificity of the assay was tested against other related viruses such as Dengue (serotypes 1-4), Japanese encephalitis, Herpes Simplex, Parainfluenza, Sindbis, Ross River, Yellow fever and West Nile viruses. The sensitivity, specificity and efficiency of this assay were 100%, 100% and 96.8% respectively. This study on early diagnostics is of importance to all endemic countries, especially Malaysia, which has been facing increasingly frequent and bigger outbreaks due to this virus since 1999.
authors.affiliation: Institute for Medical Research, Medical Entomology Unit, Infectious Diseases Research Centre, Jalan Pahang, Kuala Lumpur 50588, Malaysia.
authors.forename: A
authors.initials: A
authors.lastname: Ummul Haninah
authors.forename: S S
authors.initials: SS
authors.lastname: Vasan
authors.forename: T
authors.initials: T
authors.lastname: Ravindran
authors.forename: A
authors.initials: A
authors.lastname: Chandru
authors.forename: H L
authors.initials: HL
authors.lastname: Lee
authors.forename: S
authors.initials: S
authors.lastname: Shamala Devi
mesh_terms: D018354:Alphavirus Infections / Q000662:veterinary* / Q000821:virology*; D000818:Animals; D052160:Benzothiazoles; D002646:Chikungunya virus / Q000302:isolation & purification*; D009033:Culicidae / Q000821:virology*; D003959:Diamines; D000882:Haplorhini / Q000821:virology*; D006801:Humans; D008296:Malaysia; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D014759:Viral Envelope Proteins / Q000235:genetics; D019562:Viral Load / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral; D014759:Viral Envelope Proteins; C098022:SYBR Green I