Ultrasensitive Reverse Transcriptase Loop-Mediated Isothermal Amplification (US-LAMP)-Based Detection of Malaria Infection from Dried Blood Spots

Abstract

Submicroscopic malaria diagnosis requires highly sensitive tools instead of the conventional microscopy and rapid diagnostic tests (RDTs). While polymerase chain reaction (PCR) is more sensitive than RDTs and microscopy, the required capital cost and technical expertise hinder implementation of PCR in low- and middle-income countries. This chapter describes an ultrasensitive reverse transcriptase loop-mediated isothermal amplification (US-LAMP) test for malaria with a high sensitivity and specificity, while also being practical to implement in low-complexity laboratory settings. The workflow combines a silica spin column-based total nucleic extraction from dried blood spots (DBS) with US-LAMP amplifying the Plasmodium (Pan-LAMP) target and subsequent identification Plasmodium falciparum (Pf-LAMP).

Keywords: Asymptomatic malaria; Dried blood spots; LAMP; Malaria; Plasmodium; US-LAMP; Ultrasensitive.