A new visually improved and sensitive loop mediated isothermal amplification (LAMP) for diagnosis of symptomatic falciparum malaria

Abu Naser Mohon, Rubayet Elahi, Wasif A. Khan, Rashidul Haque, David J Sullivan, Mohammad Shafiul Alam

Research output: Contribution to journalArticle

Abstract

Molecular diagnosis of malaria by nucleotide amplification requires sophisticated and expensive instruments, typically found only in well-established laboratories. Loop-mediated isothermal amplification (LAMP) has provided a new platform for an easily adaptable molecular technique for molecular diagnosis of malaria without the use of expensive instruments. A new primer set has been designed targeting the 18S rRNA gene for the detection of Plasmodium falciparum in whole blood samples. The efficacy of LAMP using the new primer set was assessed in this study in comparison to that of a previously described set of LAMP primers as well as with microscopy and real-time PCR as reference methods for detecting P. falciparum. Pre-addition of hydroxy napthol blue (HNB) in the LAMP reaction caused a distinct color change, thereby improving the visual detection system. The new LAMP assay was found to be 99.1% sensitive compared to microscopy and 98.1% when compared to real-time PCR. Meanwhile, its specificity was 99% and 100% in contrast to microscopy and real-time PCR, respectively. Moreover, the LAMP method was in very good agreement with microscopy and real-time PCR (0.94 and 0.98, respectively). This new LAMP method can detect at least 5. parasites/μL of infected blood within 35. min, while the other LAMP method tested in this study, could detect a minimum of 100. parasites/μL of human blood after 60. min of amplification. Thus, the new method is sensitive and specific, can be carried out in a very short time, and can substitute PCR in healthcare clinics and standard laboratories.

Original languageEnglish (US)
Pages (from-to)52-57
Number of pages6
JournalActa Tropica
Volume134
Issue number1
DOIs
StatePublished - 2014
Externally publishedYes

Fingerprint

Falciparum Malaria
malaria
Real-Time Polymerase Chain Reaction
Microscopy
microscopy
Plasmodium falciparum
quantitative polymerase chain reaction
Malaria
Parasites
blood
rRNA Genes
methodology
parasites
Nucleotides
Color
loop-mediated isothermal amplification
Delivery of Health Care
Polymerase Chain Reaction
health services
nucleotides

Keywords

  • Bangladesh
  • Hydroxy napthol blue
  • LAMP
  • Malaria
  • Plasmodium falciparum

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases
  • Medicine(all)

Cite this

A new visually improved and sensitive loop mediated isothermal amplification (LAMP) for diagnosis of symptomatic falciparum malaria. / Mohon, Abu Naser; Elahi, Rubayet; Khan, Wasif A.; Haque, Rashidul; Sullivan, David J; Alam, Mohammad Shafiul.

In: Acta Tropica, Vol. 134, No. 1, 2014, p. 52-57.

Research output: Contribution to journalArticle

Mohon, Abu Naser ; Elahi, Rubayet ; Khan, Wasif A. ; Haque, Rashidul ; Sullivan, David J ; Alam, Mohammad Shafiul. / A new visually improved and sensitive loop mediated isothermal amplification (LAMP) for diagnosis of symptomatic falciparum malaria. In: Acta Tropica. 2014 ; Vol. 134, No. 1. pp. 52-57.
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