title: Comparison of real time polymerase chain reaction with microscopy and antigen detection assay for the diagnosis of malaria.
abstract: OBJECTIVE
  To determine the sensitivity of a real time polymerase chain reaction (PCR) for malaria diagnosis and to compare its accuracy with microscopy and an antigen based rapid diagnostic test (OptiMal).
  
  
  METHODS
  Cross-sectional analytical study.
  
  
  METHODS
  Military Hospital, Armed Forces Institute of Transfusion and Armed Forces Institute of Pathology, Rawalpindi, from July to December 2011.
  
  
  METHODS
  Venous blood samples of 300 clinically suspected patients of malaria were tested for malaria parasite by microscopy and OptiMal; and malaria parasite index was calculated for the positive samples. Plasmodium genus specific real time PCR was performed on all specimens, targeting small subunit rRNA gene. Diagnostic accuracy of three tests was compared and cost analysis was done.
  
  
  RESULTS
  Out of 300 patients, malaria parasite was detected in 110, 106 and 123 patients by microscopy, OptiMAL and PCR respectively. Real time PCR was 100% sensitive while microscopy and OptiMal had sensitivity of 89.4% and 86.2% respectively. All methods were 100% specific. The cost per test was calculated to be 0.2, 2.75 and 3.30 US$ by microscopy, OptiMal and PCR respectively, excluding the once capital cost on PCR equipment.
  
  
  CONCLUSIONS
  Genus specific real time PCR for the diagnosis of malaria was successfully established as a highly sensitive and affordable technology that should be incorporated in the diagnostic algorithm in this country.
authors.affiliation: Department of Haematology, Armed Forces Institute of Pathology, Rawalpindi.
authors.forename: Saleem Ahmed
authors.initials: SA
authors.lastname: Khan
authors.forename: Suhaib
authors.initials: S
authors.lastname: Ahmed
authors.forename: Nuzhat
authors.initials: N
authors.lastname: Mushahid
authors.forename: Masood
authors.initials: M
authors.lastname: Anwer
authors.forename: Shahzad
authors.initials: S
authors.lastname: Saeed
authors.forename: Farooq Ahmed
authors.initials: FA
authors.lastname: Khan
authors.forename: Ghassan Umair
authors.initials: GU
authors.lastname: Shamshad
authors.forename: Zulqarnain
authors.initials: Z
authors.lastname: Joyia
mesh_terms: D000293:Adolescent; D003365:Costs and Cost Analysis; D003430:Cross-Sectional Studies; D003955:Diagnostic Tests, Routine / Q000191:economics / Q000379:methods*; D005260:Female; D020459:Genes, rRNA / Q000235:genetics; D006801:Humans; D008288:Malaria / Q000097:blood / Q000175:diagnosis* / Q000469:parasitology; D008297:Male; D008853:Microscopy / Q000191:economics / Q000379:methods*; D010961:Plasmodium / Q000235:genetics / Q000302:isolation & purification* / Q000469:parasitology; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors