Send to

Choose Destination
Parasit Vectors. 2016 May 13;9(1):281. doi: 10.1186/s13071-016-1572-8.

Mobile suitcase laboratory for rapid detection of Leishmania donovani using recombinase polymerase amplification assay.

Author information

Center for Nutrition and Food Security, International Center for Diarrheal Disease Research, Bangladesh, Dhaka, Bangladesh.
Division of Microbiology and Animal Hygiene, Georg-August-University, Goettingen, Germany.
Department of Microbiology and Immunology, McGill University, Montréal, QC, Canada.
University Medical Centre Freiburg, Centre for Medicine and Society, Freiburg, Germany.
UNICEF/UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases (TDR), Geneva, Switzerland.
Centre for Tropical Medicine and Global Health, University of Oxford, Oxford, UK.
Division of Microbiology and Animal Hygiene, Georg-August-University, Goettingen, Germany.



Leishmania donovani (LD) is a protozoan parasite transmitted to humans from sand flies, which causes Visceral Leishmaniasis (VL). Currently, the diagnosis is based on presence of the anti-LD antibodies and clinical symptoms. Molecular diagnosis would require real-time PCR, which is not easy to implement at field settings. In this study, we report on the development and testing of a recombinase polymerase amplification (RPA) assay for the detection of LD.


A genomic DNA sample was applied to determine the assay analytical sensitivity. The cross-reactivity of the assay was tested by DNA of Leishmania spp. and of pathogens considered for differential diagnosis. The clinical performance of the assay was evaluated on LD positive and negative samples. All results were compared with real-time PCR. To allow the use of the assay at field settings, a mobile suitcase laboratory (56 × 45.5 × 26.5 cm) was developed and operated at the local hospital in Mymensingh, Bangladesh.


The LD RPA assay detected equivalent to one LD genomic DNA. The assay was performed at constant temperature (42 °C) in 15 min. The RPA assay also detected other Leishmania species (L. major, L. aethiopica and L. infantum), but did not identify nucleic acid of other pathogens. Forty-eight samples from VL, asymptomatic and post-kala-azar dermal leishmaniasis subjects were detected positive and 48 LD-negative samples were negative by both LD RPA and real-time PCR assays, which indicates 100 % agreement. The suitcase laboratory was successfully operated at the local hospital by using a solar-powered battery. DNA extraction was performed by a novel magnetic bead based method (SpeedXtract), in which a simple fast lysis protocol was applied. Moreover, All reagents were cold-chain independent.


The mobile suitcase laboratory using RPA is ideal for rapid sensitive and specific detection of LD especially at low resource settings and could contribute to VL control and elimination programmes.


Bangladesh; Leishmania donovani; Recombinase polymerase amplification assay; Suitcase laboratory; Visceral leishmaniasis

[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for BioMed Central Icon for PubMed Central
Loading ...
Support Center