Field evaluation of the photo-induced electron transfer fluorogenic primers (PET) real-time PCR for the detection of Plasmodium falciparum in Tanzania

Details

• Alternative Title:
  Malar J
• Personal Author:
  Talundzic, Eldin ; Maganga, Mussa ; Masanja, Irene M ; Peterson, David S ; Udhayakumar, Venkatachalam ; Lucchi, Naomi W
• Description:
  Background
  
  Accurate diagnosis of malaria infections remains challenging, especially in the identification of submicroscopic infections. New molecular diagnostic tools that are inexpensive, sensitive enough to detect low-level infections and suitable in laboratory settings of resource-limited countries are required for malaria control and elimination programmes. Here the diagnostic potential of a recently developed photo-induced electron transfer fluorogenic primer (PET) real-time polymerase chain reaction (PCR) called PET-PCR was investigated. This study aimed to (i) evaluate the use of this assay as a method for the detection of both Plasmodium falciparum and other Plasmodium species infections in a developing country’s diagnostic laboratory; and, (ii) determine the assay’s sensitivity and specificity compared to a nested 18S rRNA PCR.
  
  Methods
  
  Samples used in this study were obtained from a previous study conducted in the region of Iringa, Tanzania. A total of 303 samples from eight health facilities in Tanzania were utilized for this evaluation. All samples were screened using the multiplex PET-PCR assay designed to detect Plasmodium genus and P. falciparum initially in laboratory in Tanzania and then repeated at a reference laboratory at the CDC in the USA. Microscopy data was available for all the 303 samples. A subset of the samples were tested in a blinded fashion to find the sensitivity and specificity of the PET-PCR compared to the nested 18S rRNA PCR.
  
  Results
  
  Compared to microscopy, the PET-PCR assay was 59% more sensitive in detecting P. falciparum infections. The observed sensitivity and specificity were 100% (95% confidence interval (CI0.95) = 94-100%) and (CI0.95 = 96-100%), respectively, for the PET-PCR assay when compared to nested 18S rRNA PCR. When compared to 18S rRNA PCR, microscopy had a low sensitivity of 40% (CI0.95 = 23-61%) and specificity of 100% (CI0.95 = 96-100%). The PET-PCR results performed in the field laboratory in Tanzania were in 100% concordance with the results obtained at the reference laboratory in the USA.
  
  Conclusion
  
  The PET-PCR is a new molecular diagnostic tool with similar performance characteristics as commonly used PCR methods that is less expensive, easy to use, and amiable to large scale-surveillance studies in developing country settings.
  
  
• Subjects:
  DNA, Protozoan DNA Primers Fluorescent Dyes Malaria, Falciparum Microscopy Molecular Diagnostic Techniques Multiplex Polymerase Chain Reaction Plasmodium Falciparum Real-Time Polymerase Chain Reaction Research RNA, Ribosomal, 18S Sensitivity And Specificity

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• Source:
  Malar J. 2014; 13:31.
• Document Type:
  Journal Article
• Funding:
  T32 AI060546/AI/NIAID NIH HHS/United States
• Place as Subject:
  Tanzania
• Volume:
  13
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:44a5fe1aafbb23e7b0eaf36fe8721e314f16103aad421f1e800bfd1b9b545048
• Download URL:
  https://stacks.cdc.gov/view/cdc/22371/cdc_22371_DS1.pdf
• File Type:
  [PDF - 808.80 KB ]