title: Utility of multiplex reverse transcriptase-polymerase chain reaction for diagnosis and serotypic characterization of dengue and chikungunya viruses in clinical samples.
abstract: The reemergence of chikungunya virus (CHIKV) has compounded the already existing dengue problem because of clinical similarities and common vector, demanding the need for a rapid and specific diagnosis. Thus, dengue chikungunya multiplex reverse transcriptase-polymerase chain reaction (DCmRT-PCR) was developed and validated for simultaneous detection of dengue and chikungunya viral infections and its utility in virus serotyping. Blood samples from 97 suspected dengue and chikungunya cases and 10 healthy controls were subjected to dengue and chikungunya conventional RT-PCR and DCmRT-PCR. Thirty-one of 97 samples were positive for dengue or chikungunya viral RNA by RT-PCR and DCmRT-PCR with 100% concordance. DCmRT-PCR products were cycle sequenced. Seven dengue virus strains were clustered within genotype III of DENV-3 and 4 within genotype III of DENV-1, whereas chikungunya sequences were clustered within the Central/East African genotype. DCmRT-PCR was found to be a potential rapid test for simultaneous detection of dengue and CHIKV in clinical samples along with dengue serotyping.
authors.affiliation: Department of Virology, Post Graduate Institute of Medical Education and Research, Chandigarh, India. bm_mishra@hotmail.com
authors.forename: Baijayantimala
authors.initials: B
authors.lastname: Mishra
authors.forename: Mirnalini
authors.initials: M
authors.lastname: Sharma
authors.forename: Sujit Kumar
authors.initials: SK
authors.lastname: Pujhari
authors.forename: Radha Kanta
authors.initials: RK
authors.lastname: Ratho
authors.forename: Dvr Sai
authors.initials: DS
authors.lastname: Gopal
authors.forename: Cvm Naresh
authors.initials: CN
authors.lastname: Kumar
authors.forename: Gita
authors.initials: G
authors.lastname: Sarangi
authors.forename: Nirupama
authors.initials: N
authors.lastname: Chayani
authors.forename: Subhash C
authors.initials: SC
authors.lastname: Varma
mesh_terms: D000330:Aedes; D018354:Alphavirus Infections / Q000175:diagnosis* / Q000821:virology; D000818:Animals; D002460:Cell Line; D065632:Chikungunya Fever; D002646:Chikungunya virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D003715:Dengue / Q000175:diagnosis* / Q000821:virology; D003716:Dengue Virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007194:India; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017423:Sequence Analysis, RNA; D012703:Serotyping
chemical_list: D012367:RNA, Viral