title: Evaluation of non-instrumented nucleic acid amplification by loop-mediated isothermal amplification (NINA-LAMP) for the diagnosis of malaria in Northwest Ethiopia.
abstract: BACKGROUND
  Malaria is a major public health problem in sub-Saharan African countries including Ethiopia. Early and accurate diagnosis followed by prompt and effective treatment is among the various tools available for prevention, control and elimination of malaria. This study aimed to evaluate the performance of non-instrumented nucleic acid amplification loop-mediated isothermal amplification (NINA-LAMP) compared to standard thick and thin film microscopy and nested PCR as gold standard for the sensitive diagnosis of malaria in Northwest Ethiopia.
  
  
  METHODS
  A cross-sectional study was conducted in North Gondar, Ethiopia from March to July 2014. Eighty-two blood samples were collected from malaria suspected patients visiting Kola Diba Health Centre and analysed for Plasmodium parasites by microscopy, NINA-LAMP and nested PCR. The NINA-LAMP method was performed using the Loopamp Malaria Pan/Pf detection kits for detecting DNA of the genus Plasmodium and more specifically Plasmodium falciparum using an electricity-free heater. Diagnostic accuracy outcome measures (analytical sensitivity, specificity, predictive values, and Kappa scores) of NINA-LAMP and microscopy were compared to nested PCR.
  
  
  RESULTS
  A total of 82 samples were tested in the primary analysis. Using nested PCR as reference, the sensitivity and specificity of the primary NINA-LAMP assay were 96.8% (95% confidence interval (CI), 83.2% - 99.5%) and 84.3% (95% CI, 71.4% - 92.9%), respectively for detection of Plasmodium genus, and 100% (95% CI, 75.1% - 100%) and 81.2% (95% CI, 69.9% - 89.6%), respectively for detection of P. falciparum parasite. Microscopy demonstrated sensitivity and specificity of 93.6% (95% CI, 78.5% - 99.0%) and 98.0% (95% CI, 89.5% - 99.7%), respectively for the detection of Plasmodium parasites. Post-hoc repeat NINA-LAMP analysis showed improvement in diagnostic accuracy, which was comparable to nested PCR performance and superior to microscopy for detection at both the Plasmodium genus level and P. falciparum parasites.
  
  
  CONCLUSIONS
  NINA-LAMP is highly sensitive for the diagnosis of malaria and detection of Plasmodium parasite infection at both the genus and species level when compared to nested PCR. NINA-LAMP is more sensitive than microscopy for the detection of P. falciparum and differentiation from non-falciparum species and may be a critical diagnostic modality in efforts to eradicate malaria from areas of low endemicity.
authors.affiliation: Department of Medical Laboratory Sciences, College of Medicine and Health Sciences, Wollo University, Dessie, Ethiopia. mesem2000@gmail.com.
authors.forename: Meslo
authors.initials: M
authors.lastname: Sema
authors.affiliation: School of Medicine, College of Health Sciences and Medicine, Wolaita Sodo University, Wolaita, Ethiopia. yanbule@gmail.com.
authors.forename: Abebe
authors.initials: A
authors.lastname: Alemu
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Calgary, Calgary, Canada. agbayih@ucalgary.ca.
authors.forename: Abebe Genetu
authors.initials: AG
authors.lastname: Bayih
authors.affiliation: Department of Medical Parasitology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia. sisaygetie@yahoo.com.
authors.forename: Sisay
authors.initials: S
authors.lastname: Getie
authors.affiliation: Department of Medical Parasitology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia. ggebeyaw@yahoo.com.
authors.forename: Gebeyaw
authors.initials: G
authors.lastname: Getnet
authors.affiliation: PATH, Seattle, USA. dguelig@path.org.
authors.forename: Dylan
authors.initials: D
authors.lastname: Guelig
authors.affiliation: PATH, Seattle, USA. rburton@path.org.
authors.forename: Robert
authors.initials: R
authors.lastname: Burton
authors.affiliation: PATH, Seattle, USA. plabarre@path.org.
authors.forename: Paul
authors.initials: P
authors.lastname: LaBarre
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Calgary, Calgary, Canada. drpillai@ucalgary.ca.|Department of Medical Parasitology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia. drpillai@ucalgary.ca.
authors.forename: Dylan R
authors.initials: DR
authors.lastname: Pillai
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D003430:Cross-Sectional Studies; D005002:Ethiopia; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008288:Malaria / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D055815:Young Adult