Research Article: Development and clinical performance of high throughput loop-mediated isothermal amplification for detection of malaria

Date Published: February 6, 2017

Publisher: Public Library of Science

Author(s): Rushini S. Perera, Xavier C. Ding, Frank Tully, James Oliver, Nigel Bright, David Bell, Peter L. Chiodini, Iveth J. Gonzalez, Spencer D. Polley, Georges Snounou.


Accurate and efficient detection of sub-microscopic malaria infections is crucial for enabling rapid treatment and interruption of transmission. Commercially available malaria LAMP kits have excellent diagnostic performance, though throughput is limited by the need to prepare samples individually. Here, we evaluate the clinical performance of a newly developed high throughput (HTP) sample processing system for use in conjunction with the Eiken malaria LAMP kit.

The HTP system utilised dried blood spots (DBS) and liquid whole blood (WB), with parallel sample processing of 94 samples per run. The system was evaluated using 699 samples of known infection status pre-determined by gold standard nested PCR.

The sensitivity and specificity of WB-HTP-LAMP was 98.6% (95% CI, 95.7–100), and 99.7% (95% CI, 99.2–100); sensitivity of DBS-HTP-LAMP was 97.1% (95% CI, 93.1–100), and specificity 100% against PCR. At parasite densities greater or equal to 2 parasites/μL, WB and DBS HTP-LAMP showed 100% sensitivity and specificity against PCR. At densities less than 2 p/μL, WB-HTP-LAMP sensitivity was 88.9% (95% CI, 77.1–100) and specificity was 99.7% (95% CI, 99.2–100); sensitivity and specificity of DBS-HTP-LAMP was 77.8% (95% CI, 54.3–99.5) and 100% respectively.

The HTP-LAMP system is a highly sensitive diagnostic test, with the potential to allow large scale population screening in malaria elimination campaigns.

Partial Text

Successful elimination of malaria is dependent on accurate and efficient diagnoses to actively treat and clear parasite infections, thereby interrupting transmission to new hosts. Microscopic examination of thick and thin blood smears remains a commonly used method for diagnosis of malarial infections [1], capable of detecting parasites in less than 60 minutes. Microscopy-based diagnosis is completely dependent upon the skill of trained microscopists and implementation of regular external quality assessment (EQA) schemes. The development of antigen-based rapid diagnostic tests (RDTs) allows for quick and accurate detection of infections [2]. Such testing is relied upon in areas lacking trained microscopists and specialised diagnostic equipment, and is increasingly being used for diagnostic testing over microscopy [3]. However, both techniques have limited sensitivity: the lowest detection limit is 5 to 50 parasites per microLitre (p/μL) of blood for expert microscopy, and around 100 p/μL for RDTs-, and therefore inadequate for detection of infections below these levels, which frequently occur [1, 3–6].

Active identification and treatment of sub-patent infections in asymptomatic parasite carriers is crucial to achieving the World Health Organization (WHO) goal of 90% reduction in global malaria incidence and mortality rates by 2030 [30].