title: Differential Diagnosis of Malaria on Truelab Uno®, a Portable, Real-Time, MicroPCR Device for Point-Of-Care Applications.
abstract: BACKGROUND
  Sensitive and specific detection of malarial parasites is crucial in controlling the significant malaria burden in the developing world. Also important is being able to identify life threatening Plasmodium falciparum malaria quickly and accurately to reduce malaria related mortality. Existing methods such as microscopy and rapid diagnostic tests (RDTs) have major shortcomings. Here, we describe a new real-time PCR-based diagnostic test device at point-of-care service for resource-limited settings.
  
  
  METHODS
  Truenat® Malaria, a chip-based microPCR test, was developed by bigtec Labs, Bangalore, India, for differential identification of Plasmodium falciparum and Plasmodium vivax parasites. The Truenat Malaria tests runs on bigtec's Truelab Uno® microPCR device, a handheld, battery operated, and easy-to-use real-time microPCR device. The performance of Truenat® Malaria was evaluated versus the WHO nested PCR protocol. The Truenat® Malaria was further evaluated in a triple-blinded study design using a sample panel of 281 specimens created from the clinical samples characterized by expert microscopy and a rapid diagnostic test kit by the National Institute of Malaria Research (NIMR). A comparative evaluation was done on the Truelab Uno® and a commercial real-time PCR system.
  
  
  RESULTS
  The limit of detection of the Truenat Malaria assay was found to be <5 parasites/μl for both P. falciparum and P. vivax. The Truenat® Malaria test was found to have sensitivity and specificity of 100% each, compared to the WHO nested PCR protocol based on the evaluation of 100 samples. The sensitivity using expert microscopy as the reference standard was determined to be around 99.3% (95% CI: 95.5-99.9) at the species level. Mixed infections were identified more accurately by Truenat Malaria (32 samples identified as mixed) versus expert microscopy and RDTs which detected 4 and 5 mixed samples, respectively.
  
  
  CONCLUSIONS
  The Truenat® Malaria microPCR test is a valuable diagnostic tool and implementation should be considered not only for malaria diagnosis but also for active surveillance and epidemiological intervention.
authors.affiliation: bigtec Private Limited, 2nd Floor, Golden Heights, 59th C Cross, 4th M Block, Rajajinagar, Bangalore, 560 010, Karnataka, India.|School of Bio Sciences and Technology, School of Advanced Sciences VIT University, Vellore, Tamil Nadu, 632014, India.
authors.forename: Chandrasekhar Bhaskaran
authors.initials: CB
authors.lastname: Nair
authors.affiliation: bigtec Private Limited, 2nd Floor, Golden Heights, 59th C Cross, 4th M Block, Rajajinagar, Bangalore, 560 010, Karnataka, India.
authors.forename: Jagannath
authors.initials: J
authors.lastname: Manjula
authors.affiliation: National Institute of Malaria Research, ICMR Complex, Poojanahalli, Kannamangala Post, Devanahalli, Bangalore, 562 110, Karnataka, India.
authors.forename: Pradeep Annamalai
authors.initials: PA
authors.lastname: Subramani
authors.affiliation: Institute of Trans-disciplinary Health Sciences and Technology (IHST), FRLHT, 74/2, Jarakabande Kaval, Attur post, Via Yelahanka, Bangalore, 560 106, Karnataka, India.
authors.forename: Prakash B
authors.initials: PB
authors.lastname: Nagendrappa
authors.affiliation: bigtec Private Limited, 2nd Floor, Golden Heights, 59th C Cross, 4th M Block, Rajajinagar, Bangalore, 560 010, Karnataka, India.
authors.forename: Mulakkapurath Narayanan
authors.initials: MN
authors.lastname: Manoj
authors.affiliation: bigtec Private Limited, 2nd Floor, Golden Heights, 59th C Cross, 4th M Block, Rajajinagar, Bangalore, 560 010, Karnataka, India.
authors.forename: Sukriti
authors.initials: S
authors.lastname: Malpani
authors.affiliation: bigtec Private Limited, 2nd Floor, Golden Heights, 59th C Cross, 4th M Block, Rajajinagar, Bangalore, 560 010, Karnataka, India.
authors.forename: Phani Kumar
authors.initials: PK
authors.lastname: Pullela
authors.affiliation: bigtec Private Limited, 2nd Floor, Golden Heights, 59th C Cross, 4th M Block, Rajajinagar, Bangalore, 560 010, Karnataka, India.
authors.forename: Pillarisetti Venkata
authors.initials: PV
authors.lastname: Subbarao
authors.affiliation: School of Bio Sciences and Technology, School of Advanced Sciences VIT University, Vellore, Tamil Nadu, 632014, India.
authors.forename: Siva
authors.initials: S
authors.lastname: Ramamoorthy
authors.affiliation: National Institute of Malaria Research, ICMR Complex, Poojanahalli, Kannamangala Post, Devanahalli, Bangalore, 562 110, Karnataka, India.
authors.forename: Susanta K
authors.initials: SK
authors.lastname: Ghosh
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D003430:Cross-Sectional Studies; D003937:Diagnosis, Differential; D003955:Diagnostic Tests, Routine / Q000295:instrumentation*; D042241:Early Diagnosis; D006801:Humans; D007194:India; D016778:Malaria, Falciparum / Q000175:diagnosis* / Q000469:parasitology; D016780:Malaria, Vivax / Q000175:diagnosis* / Q000469:parasitology; D008875:Middle Aged; D010963:Plasmodium falciparum / Q000235:genetics*; D010966:Plasmodium vivax / Q000235:genetics*; D019095:Point-of-Care Systems; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D012680:Sensitivity and Specificity; D055815:Young Adult