title: Nested reverse transcription polymerase chain reaction for the detection of rubella virus in clinical specimens.
abstract: OBJECTIVE
  Rubella virus (RV) is one of the leading causes of childhood blindness in India. In this study we applied an optimized nested reverse transcription polymerase chain reaction (nRT-PCR) to detect RV in clinical specimens.
  
  
  METHODS
  nRT-PCR was optimized using total RNA extracted from standard strain of RV using nested sets of primers specific for E1 open reading frame. nRT-PCR was applied onto 30 lens aspirates and corresponding peripheral blood leucocytes of 30 infants with congenital (29)/ developmental (01) cataract. Serology for anti-RV IgG and IgM antibodies was done. RV isolation was attempted using Vero and SIRC cell cultures.
  
  
  RESULTS
  Optimized nRT-PCR was specific for RV and sensitive to detect 10 fg of RV RNA. Among 30 patients, nRT-PCR detected presence of RV in lens aspirates of 6 (20%) and 4 corresponding leucocytes. RV was isolated from 3 (10%) lens aspirates (nRT-PCR positive) of the 30 patients. Sera of these 6 patients showed presence of anti-RV IgG and IgM in one, only anti-RV IgG in 3 others and none in the other two. Of the remaining 24 patients, anti-RV IgG was detected in 3 and no anti-RV IgM antibodies in others.
  
  
  CONCLUSIONS
  Findings of our study showed that the nRT-PCR was a more sensitive and rapid technique to detect RV from lens aspirates compared to conventional methods of virus isolation and serology.
authors.affiliation: L & T Microbiology Research Centre, Vision Research Foundation, Sankara Nethralaya, Chennai, India.
authors.forename: G
authors.initials: G
authors.lastname: Shyamala
authors.forename: J
authors.initials: J
authors.lastname: Malathi
authors.forename: Y S
authors.initials: YS
authors.lastname: Moses
authors.forename: K L
authors.initials: KL
authors.lastname: Therese
authors.forename: H N
authors.initials: HN
authors.lastname: Madhavan
mesh_terms: D000818:Animals; D002386:Cataract / Q000151:congenital / Q000821:virology; D002522:Chlorocebus aethiops; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007908:Lens, Crystalline / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012409:Rubella / Q000151:congenital / Q000821:virology; D012412:Rubella virus / Q000235:genetics / Q000302:isolation & purification*; D014709:Vero Cells
chemical_list: D012367:RNA, Viral