title: Simple, rapid, inexpensive platform for the diagnosis of malaria by loop mediated isothermal amplification (LAMP).
abstract: We attempted to improve the loop-mediated isothermal amplification (LAMP) method for malaria diagnosis by using a simple DNA extraction procedure, and a portable device performing both the amplification and detection of LAMP in one platform. Additionally, the device served as a heating block for the DNA preparation. We refer this method as LAMP-Tube scanner, and evaluated using 209 microscopically positive malaria samples and compared them to RDTs and LAMP-Thermocycler. Two most common human infecting Plasmodium species were detected. The LAMP-Tube scanner method is found to be simple and allowed real-time detection of DNA amplification. The time to amplification varied but was closely less than 60 min. Sensitivity and specificity of LAMP-Tube scanner in detecting Plasmodium falciparum were 95% and 93.3%, compared to microscopy and 98.3% and 100% respectively, compared to standard LAMP-Thermocycler. In addition, it showed a detection limit of 10 and 40 copies of the parasitemia for Plasmodium vivax and P. falciparum. Accordingly, in comparison to the results obtained by microscopy, the LAMP-Tube scanner had a less divergence in sensitivity and specificity, and yielded results similar to those of LAMP-Thermocycler. This method has the great potential as a field usable molecular tool for the diagnosis of malaria and is an alternative to conventional PCR-based diagnostic methods for field use.
authors.affiliation: Genomix Molecular Diagnostics (P) Ltd., 5-36/207 Prasanthnagar, Kukatpally, Hyderabad 500 072, India. ramsura@gmail.com
authors.forename: Rambabu
authors.initials: R
authors.lastname: Surabattula
authors.forename: Manju Pradeep
authors.initials: MP
authors.lastname: Vejandla
authors.forename: Prudhvi Chand
authors.initials: PC
authors.lastname: Mallepaddi
authors.forename: Konrad
authors.initials: K
authors.lastname: Faulstich
authors.forename: Rathnagiri
authors.initials: R
authors.lastname: Polavarapu
mesh_terms: D016054:DNA, Protozoan / Q000097:blood / Q000302:isolation & purification*; D006801:Humans; D008288:Malaria / Q000175:diagnosis* / Q000469:parasitology; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D010963:Plasmodium falciparum / Q000235:genetics*; D010966:Plasmodium vivax / Q000235:genetics*; D019095:Point-of-Care Systems; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D016054:DNA, Protozoan
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