Loop Mediated Isothermal Amplification (LAMP) Accurately Detects Malaria DNA from Filter Paper Blood Samples of Low Density Parasitaemias

Aydin-Schmidt, Berit; Xu, Weiping; González, Iveth J.; Polley, Spencer D.; Bell, David; Shakely, Delér; Msellem, Mwinyi I.; Björkman, Anders; Mårtensson, Andreas
August 2014
PLoS ONE;Aug2014, Vol. 9 Issue 8, p1
Academic Journal
Background: Loop mediated isothermal amplification (LAMP) provides an opportunity for improved, field-friendly detection of malaria infections in endemic areas. However data on the diagnostic accuracy of LAMP for active case detection, particularly low-density parasitaemias, are lacking. We therefore evaluated the performance of a new LAMP kit compared with PCR using DNA from filter paper blood spots. Methods and Findings: Samples from 865 fever patients and 465 asymptomatic individuals collected in Zanzibar were analysed for Pan (all species) and Pf (P. falciparum) DNA with the Loopamp MALARIA Pan/Pf kit. Samples were amplified at 65°C for 40 minutes in a real-time turbidimeter and results were compared with nested PCR. Samples with discordant results between LAMP and nested PCR were analysed with real-time PCR. The real-time PCR corrected nested PCR result was defined as gold standard. Among the 117 (13.5%) PCR detected P. falciparum infections from fever patients (mean parasite density 7491/µL, range 6–782,400) 115, 115 and 111 were positive by Pan-LAMP, Pf-LAMP and nested PCR, respectively. The sensitivities were 98.3% (95%CI 94–99.8) for both Pan and Pf-LAMP. Among the 54 (11.6%) PCR positive samples from asymptomatic individuals (mean parasite density 10/µL, range 0–4972) Pf-LAMP had a sensitivity of 92.7% (95%CI 80.1–98.5) for detection of the 41 P. falciparum infections. Pan-LAMP had sensitivities of 97% (95%CI 84.2–99.9) and 76.9% (95%CI 46.2–95) for detection of P. falciparum and P. malariae, respectively. The specificities for both Pan and Pf-LAMP were 100% (95%CI 99.1–100) in both study groups. Conclusion: Both components of the Loopamp MALARIA Pan/Pf detection kit revealed high diagnostic accuracy for parasite detection among fever patients and importantly also among asymptomatic individuals of low parasite densities from minute blood volumes preserved on filter paper. These data support LAMPs potential role for improved detection of low-density malaria infections in pre-elimination settings.


Related Articles

  • Comparison of three PCR-based assays for the non-invasive diagnosis of malaria: detection of Plasmodium parasites in blood and saliva. Singh, R.; Singh, D.; Gupta, R.; Savargaonkar, D.; Singh, O.; Nanda, N.; Bhatt, R.; Valecha, N. // European Journal of Clinical Microbiology & Infectious Diseases;Sep2014, Vol. 33 Issue 9, p1631 

    The conventional molecular diagnosis of malaria uses 18S rRNA-based PCR assay employing blood samples. This assay presents limitation in terms of long turnaround time and increased chances of false-positive results. Here, we evaluated one-step singleplex or multiplex PCR assay based on high copy...

  • Association of sub-microscopic malaria parasite carriage with transmission intensity in northeastern Tanzania.  // Malaria Journal;2011, Vol. 10 Issue 1, p370 

    The article presents a study that investigated the association of association of sub-microscopic malaria parasite with clonal complexity of infections in northeastern Tanzania. In the study 1,121 blood samples collected from 13 villages and analyzed by polymerase chain reaction (PCR) and then...

  • Real-Time Fluorescence Loop Mediated Isothermal Amplification for the Diagnosis of Malaria. Lucchi, Naomi W.; Demas, Allison; Narayanan, Jothikumar; Sumari, Deborah; Kabanywanyi, Abdunoor; Kachur, S. Patrick; Barnwell, John W.; Udhayakumar, Venkatachalam // PLoS ONE;2010, Vol. 5 Issue 10, p1 

    Background: Molecular diagnostic methods can complement existing tools to improve the diagnosis of malaria. However, they require good laboratory infrastructure thereby restricting their use to reference laboratories and research studies. Therefore, adopting molecular tools for routine use in...

  • Multiplex qPCR for Detection and Absolute Quantification of Malaria. Kamau, Edwin; Alemayehu, Saba; Feghali, Karla C.; Saunders, David; Ockenhouse, Christian F. // PLoS ONE;Aug2013, Vol. 8 Issue 8, p1 

    We describe development of an absolute multiplex quantitative real-time PCR for detection of Plasmodium spp., P. falciparum and P. vivax targets in order to produce an assay amenable to high throughput but with reduced costs. Important qPCR experimental details and information that is critical...

  • A Modified RNA-Seq Approach for Whole Genome Sequencing of RNA Viruses from Faecal and Blood Samples. Batty, Elizabeth M.; Wong, T. H. Nicholas; Trebes, Amy; Argoud, Karène; Attar, Moustafa; Buck, David; Ip, Camilla L. C.; Golubchik, Tanya; Cule, Madeleine; Bowden, Rory; Manganis, Charis; Klenerman, Paul; Barnes, Eleanor; Walker, A. Sarah; Wyllie, David H.; Wilson, Daniel J.; Dingle, Kate E.; Peto, Tim E. A.; Crook, Derrick W.; Piazza, Paolo // PLoS ONE;Jun2013, Vol. 8 Issue 6, p1 

    To date, very large scale sequencing of many clinically important RNA viruses has been complicated by their high population molecular variation, which creates challenges for polymerase chain reaction and sequencing primer design. Many RNA viruses are also difficult or currently not possible to...

  • Accuracy in amplification. Stenman, Jakob; Orpana, Arto // Nature Biotechnology;Nov2001, Vol. 19 Issue 11, p1011 

    Discusses the application of polymerase chain reaction (PCR) techniques. Accuracy of amplification techniques; Sensitivity of PCR; Stochastic variation in low-copy-number target templates.

  • Molecular Diagnostic Field Test for Point-of-Care Detection of Ebola Virus Directly From Blood. Benzine, Jason W.; Brown, Kerry M.; Agans, Krystle N.; Godiska, Ronald; Mire, Chad E.; Gowda, Krishne; Converse, Brandon; Geisbert, Thomas W.; Mead, David A.; Chander, Yogesh // Journal of Infectious Diseases;2016 Supplement, Vol. 214, pS234 

    A molecular diagnostic method for robust detection of Ebola virus (EBOV) at the point of care (POC) directly from blood samples is described. This assay is based on reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) of the glycoprotein gene of EBOV. Complete reaction...

  • Hypersentitive PCR, Ancient Human and mtDNA, and Contaminaton. Yang, Dongya Y.; Eng, Barry; Saunders, Shelley R. // Human Biology;Jun2003, Vol. 75 Issue 3, p355 

    Studies the amplification of human mitochondrial DNA. Polymerase chain reaction (PCR) amplification; Positive amplifications of negative controls; Sequence of PCR products; Levels of contamination; Sensitivity of PCR amplification.

  • The PCR assay in the preclinical safety evaluation of nucleic acid medicines. Haworth, R.; Pilling, A.M. // Human & Experimental Toxicology;May2000, Vol. 19 Issue 5, p267 

    The polymerase chain reaction (PCR) is a highly efficient gene amplification procedure which is increasingly being applied to the safety assessment of nucleic acid (NA) medicines such as gene therapies and DNA vaccines. Although clinical experience is limited, a number of potential safety issues...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics