title: Clinical applicability of single-tube multiplex reverse-transcriptase PCR in dengue virus diagnosis and serotyping.
abstract: This study has evaluated the clinical applicability of a single-tube multiplex RT-PCR as compared with a two-step nested RT-PCR for the diagnosis as well as serotyping of dengue virus in patient's samples. Seventy-six acute phase blood samples collected from clinically suspected dengue patients during the 2008 outbreak were subjected to two-step nested RT-PCR and single-tube multiplex RT-PCR for dengue diagnosis and serotyping. Of the 76 samples, 17 (22.4%) were positive for dengue viral RNA. Single dengue virus infection was found in 16 cases and 1 had concurrent infection with two serotypes (3&1). Dengue serotype 3 was the predominant serotype (70.5%), followed by serotype 1 (23.5%). Single-tube multiplex PCR had concordant result with that of two-step nested RT-PCR including the one with concomitant infection. This study reveals the predominance of dengue serotype 3 in North India in addition to the co-circulation of multiple serotypes and concomitant infection. The rapid and accurate diagnostic capability of single-tube multiplex RT-PCR used in the study appears to be promising enough to be commonly used for dengue viral detection as well as serotyping.
authors: Department of Virology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. bm_mishra@hotmail.com
authors: Bajayantimala
authors: B
authors: Mishra
authors: Mirnalini
authors: M
authors: Sharma
authors: Sujit Kumar
authors: SK
authors: Pujhari
authors: Suma B
authors: SB
authors: Appannanavar
authors: Radha Kanta
authors: RK
authors: Ratho
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D003715:Dengue / Q000097:blood / Q000175:diagnosis* / Q000453:epidemiology; D003716:Dengue Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D004196:Disease Outbreaks; D005260:Female; D006801:Humans; D007194:India / Q000453:epidemiology; D007223:Infant; D008297:Male; D008875:Middle Aged; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping; D055815:Young Adult
chemical_list: D012367:RNA, Viral