title: Development of a multiplex real-time RT-PCR assay for simultaneous detection of dengue and chikungunya viruses.
abstract: Dengue and chikungunya viruses co-circulate and cause infections that start with similar symptoms but progress to radically different outcomes. Therefore, an early diagnostic test that can differentiate between the two is needed. A single-step multiplex real-time RT-PCR assay was developed that can simultaneously detect and quantitate RNA of all dengue virus (DENV) serotypes and chikungunya virus (CHIKV). The sensitivity was 100 % for DENV and 95.8 % for CHIKV, whilst the specificity was 100 % for both viruses when compared with conventional RT-PCR. The detection limit ranged from 1 to 50 plaque-forming units. The assay was successfully used for differential diagnosis of dengue and chikungunya in Pune, where the viruses co-circulate.
authors.affiliation: Dengue/Chikungunya group, National Institute of Virology, Pune, 411001, Maharashtra, India, cecilia.dayaraj@gmail.com.
authors.forename: D
authors.initials: D
authors.lastname: Cecilia
authors.forename: M
authors.initials: M
authors.lastname: Kakade
authors.forename: K
authors.initials: K
authors.lastname: Alagarasu
authors.forename: J
authors.initials: J
authors.lastname: Patil
authors.forename: A
authors.initials: A
authors.lastname: Salunke
authors.forename: D
authors.initials: D
authors.lastname: Parashar
authors.forename: P S
authors.initials: PS
authors.lastname: Shah
mesh_terms: D065632:Chikungunya Fever / Q000175:diagnosis* / Q000821:virology; D002646:Chikungunya virus / Q000302:isolation & purification*; D003715:Dengue / Q000175:diagnosis* / Q000821:virology; D003716:Dengue Virus / Q000302:isolation & purification*; D006801:Humans; D007194:India / Q000453:epidemiology; D012367:RNA, Viral / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral