1921
Volume 97, Issue 5
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Abstract.

In this study, we developed a recombinase polymerase amplification (RPA) assay for specific diagnosis of . Genomic DNA was extracted from whole blood samples using a commercial kit. With incubation at 37°C, the samples were successfully amplified within 20 minutes. The end product of RPA was further examined by loading onto agarose gel and a specific band was observed with a size of 128 bp. The RPA assay exhibited high sensitivity with limits of detection down to one copy of the plasmid. From the specificity experiments, it was demonstrated that all samples ( = 45) were positive while other spp. ( = 42) and negative samples ( = 6) were negative. Therefore, the RPA assay is a highly promising approach with the potential to be used in resource-limited settings. This assay can be further optimized for bedside and on field application.

Loading

Article metrics loading...

The graphs shown below represent data from March 2017
/content/journals/10.4269/ajtmh.17-0427
2017-08-14
2019-05-22
Loading full text...

Full text loading...

/deliver/fulltext/14761645/97/5/tpmd170427.html?itemId=/content/journals/10.4269/ajtmh.17-0427&mimeType=html&fmt=ahah

References

  1. World Health Organization, 2016. The World Malaria Report 2016. Geneva, Switzerland: World Health Organization.
  2. Singh B, Kim Sung L, Matusop A, Radhakrishnan A, Shamsul SS, Cox-Singh J, Thomas A, Conway DJ, , 2004. A large focus of naturally acquired Plasmodium knowlesi infections in human beings. Lancet 363: 10171024.[Crossref] [Google Scholar]
  3. Lee KS, Cox-Singh J, Singh B, , 2009. Morphological features and differential counts of Plasmodium knowlesi parasites in naturally acquired human infections. Malar J 8: 73.[Crossref] [Google Scholar]
  4. Imwong M, Tanomsing N, Pukrittayakamee S, Day NP, White NJ, Snounou G, , 2009. Spurious amplification of a Plasmodium vivax small-subunit RNA gene by use of primers currently used to detect P. knowlesi . J Clin Microbiol 47: 41734175.[Crossref] [Google Scholar]
  5. Lau YL, Lai MY, Anthony CA, Chang PY, Palaeya V, Fong MY, Mahmud R, , 2015. Comparison of three molecular methods for the detection and speciation of five human Plasmodium species. Am J Trop Med Hyg 92: 2833.[Crossref] [Google Scholar]
  6. Piepenburg O, Williams CH, Stemple DL, Armes NA, , 2006. DNA detection using recombination protein. PLoS Biol 4: e204.[Crossref] [Google Scholar]
  7. Teoh BT, , 2015. Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification. J Clin Microbiol 53: 830837.[Crossref] [Google Scholar]
  8. Abd El Wahed A, , 2015. Recombinase polymerase amplification assay for rapid diagnostics of dengue infection. PLoS One 10: e0129682.[Crossref] [Google Scholar]
  9. Kersting S, Rausch V, Bier FF, Nickisch-Rosenegk MV, , 2014. Rapid detection of Plasmodium falciparum with isothermal recombinase polymerase amplification and lateral flow analysis. Malar J 13: 99.[Crossref] [Google Scholar]
  10. Moorea MD, Jaykus LA, , 2017. Development of a recombinase polymerase amplification assay for detection of epidemic human noroviruses. Sci Rep 7: 40244.[Crossref] [Google Scholar]
  11. Lau YL, Fong MY, Mahmud R, Chang PY, Palaeya V, Cheong FW, Chin LC, Anthony CN, Al-Mekhlafi AM, Chen Y, , 2014. Specific, sensitive and rapid detection of human Plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP) in blood samples. Malar J 10: 197.[Crossref] [Google Scholar]
  12. Lau YL, Lai MY, Fong MY, Jelip J, Mahmud R, , 2016. Comparison of three molecular methods for the detection and speciation of five human Plasmodium species. Am J Trop Med Hyg 94: 336339.[Crossref] [Google Scholar]
  13. Yan L, Zhou J, Zheng Y, Gamson AS, Roembke BT, Nakayama S, Sintim HO, , 2014. Isothermal amplified detection of DNA and RNA. Mol Biosyst 10: 9701003.[Crossref] [Google Scholar]
http://instance.metastore.ingenta.com/content/journals/10.4269/ajtmh.17-0427
Loading
/content/journals/10.4269/ajtmh.17-0427
Loading

Data & Media loading...

  • Received : 01 Jun 2017
  • Accepted : 30 Jun 2017
  • Published online : 14 Aug 2017

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error