Skip to main content
Top
Published in: Malaria Journal 1/2013

Open Access 01-12-2013 | Research

An innovative tool for moving malaria PCR detection of parasite reservoir into the field

Authors: Lydie Canier, Nimol Khim, Saorin Kim, Vincent Sluydts, Somony Heng, Dany Dourng, Rotha Eam, Sophy Chy, Chanra Khean, Kaknika Loch, Malen Ken, Hokkean Lim, Sovannaroath Siv, Sochantha Tho, Pascal Masse-Navette, Charlotte Gryseels, Sambunny Uk, Karel Van Roey, Koen Peeters Grietens, Mao Sokny, Boukheng Thavrin, Char Meng Chuor, Vincent Deubel, Lies Durnez, Marc Coosemans, Didier Ménard

Published in: Malaria Journal | Issue 1/2013

Login to get access

Abstract

Background

To achieve the goal of malaria elimination in low transmission areas such as in Cambodia, new, inexpensive, high-throughput diagnostic tools for identifying very low parasite densities in asymptomatic carriers are required. This will enable a switch from passive to active malaria case detection in the field.

Methods

DNA extraction and real-time PCR assays were implemented in an “in-house” designed mobile laboratory allowing implementation of a robust, sensitive and rapid malaria diagnostic strategy in the field. This tool was employed in a survey organized in the context of the MalaResT project (NCT01663831).

Results

The real-time PCR screening and species identification assays were performed in the mobile laboratory between October and November 2012, in Rattanakiri Province, to screen approximately 5,000 individuals in less than four weeks and treat parasite carriers within 24–48 hours after sample collection. An average of 240 clinical samples (and 40 quality control samples) was tested every day, six/seven days per week. Some 97.7% of the results were available <24 hours after the collection. A total of 4.9% were positive for malaria. Plasmodium vivax was present in 61.1% of the positive samples, Plasmodium falciparum in 45.9%, Plasmodium malariae in 7.0% and Plasmodium ovale in 2.0%.

Conclusions

The operational success of this diagnostic set-up proved that molecular testing and subsequent treatment is logistically achievable in field settings. This will allow the detection of clusters of asymptomatic carriers and to provide useful epidemiological information. Fast results will be of great help for staff in the field to track and treat asymptomatic parasitaemic cases. The concept of the mobile laboratory could be extended to other countries for the molecular detection of malaria or other pathogens, or to culture vivax parasites, which does not support long-time delay between sample collection and culture.
Appendix
Available only for authorised users
Literature
1.
go back to reference WHO: World Malaria Report 2012. 2012, Geneva: World Health Organization WHO: World Malaria Report 2012. 2012, Geneva: World Health Organization
2.
go back to reference Cui L, Yan G, Sattabongkot J, Cao Y, Chen B, Chen X, Fan Q, Fang Q, Jongwutiwes S, Parker D, Sirichaisinthop J, Kyaw MP, Su XZ, Yang H, Yang Z, Wang B, Xu J, Zheng B, Zhong D, Zhou G: Malaria in the Greater Mekong Subregion: heterogeneity and complexity. Acta Trop. 2012, 121: 227-239. 10.1016/j.actatropica.2011.02.016.PubMedCentralCrossRefPubMed Cui L, Yan G, Sattabongkot J, Cao Y, Chen B, Chen X, Fan Q, Fang Q, Jongwutiwes S, Parker D, Sirichaisinthop J, Kyaw MP, Su XZ, Yang H, Yang Z, Wang B, Xu J, Zheng B, Zhong D, Zhou G: Malaria in the Greater Mekong Subregion: heterogeneity and complexity. Acta Trop. 2012, 121: 227-239. 10.1016/j.actatropica.2011.02.016.PubMedCentralCrossRefPubMed
3.
go back to reference Amaratunga C, Sreng S, Suon S, Phelps ES, Stepniewska K, Lim P, Zhou C, Mao S, Anderson JM, Lindegardh N, Jiang H, Song J, Su XZ, White NJ, Dondorp AM, Anderson TJ, Fay MP, Mu J, Duong S, Fairhurst RM: Artemisinin-resistant Plasmodium falciparum in Pursat province, western Cambodia: a parasite clearance rate study. Lancet Infect Dis. 2012, 12: 851-858. 10.1016/S1473-3099(12)70181-0.PubMedCentralCrossRefPubMed Amaratunga C, Sreng S, Suon S, Phelps ES, Stepniewska K, Lim P, Zhou C, Mao S, Anderson JM, Lindegardh N, Jiang H, Song J, Su XZ, White NJ, Dondorp AM, Anderson TJ, Fay MP, Mu J, Duong S, Fairhurst RM: Artemisinin-resistant Plasmodium falciparum in Pursat province, western Cambodia: a parasite clearance rate study. Lancet Infect Dis. 2012, 12: 851-858. 10.1016/S1473-3099(12)70181-0.PubMedCentralCrossRefPubMed
4.
go back to reference Dondorp AM, Nosten F, Yi P, Das D, Phyo AP, Tarning J, Lwin KM, Ariey F, Hanpithakpong W, Lee SJ, Ringwald P, Silamut K, Imwong M, Chotivanich K, Lim P, Herdman T, An SS, Yeung S, Singhasivanon P, Day NP, Lindegardh N, Socheat D, White NJ: Artemisinin resistance in Plasmodium falciparum malaria. N Engl J Med. 2009, 361: 455-467. 10.1056/NEJMoa0808859.PubMedCentralCrossRefPubMed Dondorp AM, Nosten F, Yi P, Das D, Phyo AP, Tarning J, Lwin KM, Ariey F, Hanpithakpong W, Lee SJ, Ringwald P, Silamut K, Imwong M, Chotivanich K, Lim P, Herdman T, An SS, Yeung S, Singhasivanon P, Day NP, Lindegardh N, Socheat D, White NJ: Artemisinin resistance in Plasmodium falciparum malaria. N Engl J Med. 2009, 361: 455-467. 10.1056/NEJMoa0808859.PubMedCentralCrossRefPubMed
5.
go back to reference Noedl H, Se Y, Schaecher K, Smith BL, Socheat D, Fukuda MM: Evidence of artemisinin-resistant malaria in western Cambodia. N Engl J Med. 2008, 359: 2619-2620. 10.1056/NEJMc0805011.CrossRefPubMed Noedl H, Se Y, Schaecher K, Smith BL, Socheat D, Fukuda MM: Evidence of artemisinin-resistant malaria in western Cambodia. N Engl J Med. 2008, 359: 2619-2620. 10.1056/NEJMc0805011.CrossRefPubMed
6.
go back to reference Phyo AP, Nkhoma S, Stepniewska K, Ashley EA, Nair S, McGready R, Ler Moo C, Al-Saai S, Dondorp AM, Lwin KM, Singhasivanon P, Day NP, White NJ, Anderson TJ, Nosten F: Emergence of artemisinin-resistant malaria on the western border of Thailand: a longitudinal study. Lancet. 2012, 379: 1960-1966. 10.1016/S0140-6736(12)60484-X.PubMedCentralCrossRefPubMed Phyo AP, Nkhoma S, Stepniewska K, Ashley EA, Nair S, McGready R, Ler Moo C, Al-Saai S, Dondorp AM, Lwin KM, Singhasivanon P, Day NP, White NJ, Anderson TJ, Nosten F: Emergence of artemisinin-resistant malaria on the western border of Thailand: a longitudinal study. Lancet. 2012, 379: 1960-1966. 10.1016/S0140-6736(12)60484-X.PubMedCentralCrossRefPubMed
7.
go back to reference Kyaw MP, Nyunt MH, Chit K, Aye MM, Aye KH, Aye MM, Lindegardh N, Tarning J, Imwong M, Jacob CG, Rasmussen C, Perin J, Ringwald P, Nyunt MM: Reduced susceptibility of Plasmodium falciparum to artesunate in southern Myanmar. PLoS One. 2013, 8: e57689-10.1371/journal.pone.0057689.PubMedCentralCrossRefPubMed Kyaw MP, Nyunt MH, Chit K, Aye MM, Aye KH, Aye MM, Lindegardh N, Tarning J, Imwong M, Jacob CG, Rasmussen C, Perin J, Ringwald P, Nyunt MM: Reduced susceptibility of Plasmodium falciparum to artesunate in southern Myanmar. PLoS One. 2013, 8: e57689-10.1371/journal.pone.0057689.PubMedCentralCrossRefPubMed
8.
go back to reference Tran TH, Nguyen TT, Nguyen HP, Boni MF, Ngo VT, Nguyen TN, Le HT, Cao QT, Pham VT, Phung DT, Le TL, Le TD, Merson L, Dolecek C, Stepniewska K, Ringwald P, White NJ, Farrar J, Wolbers M: In vivo susceptibility of Plasmodium falciparum to artesunate in Binh Phuoc Province Vietnam. Malar J. 2013, 11: 355- Tran TH, Nguyen TT, Nguyen HP, Boni MF, Ngo VT, Nguyen TN, Le HT, Cao QT, Pham VT, Phung DT, Le TL, Le TD, Merson L, Dolecek C, Stepniewska K, Ringwald P, White NJ, Farrar J, Wolbers M: In vivo susceptibility of Plasmodium falciparum to artesunate in Binh Phuoc Province Vietnam. Malar J. 2013, 11: 355-
9.
go back to reference Gurarie D, Karl S, Zimmerman PA, King CH, St Pierre TG, Davis TM: Mathematical modeling of malaria infection with innate and adaptive immunity in individuals and agent-based communities. PLoS One. 2011, 7: e34040-CrossRef Gurarie D, Karl S, Zimmerman PA, King CH, St Pierre TG, Davis TM: Mathematical modeling of malaria infection with innate and adaptive immunity in individuals and agent-based communities. PLoS One. 2011, 7: e34040-CrossRef
10.
go back to reference Incardona S, Vong S, Chiv L, Lim P, Nhem S, Sem R, Khim N, Doung S, Mercereau-Puijalon O, Fandeur T: Large-scale malaria survey in Cambodia: novel insights on species distribution and risk factors. Malar J. 2007, 6: 37-10.1186/1475-2875-6-37.PubMedCentralCrossRefPubMed Incardona S, Vong S, Chiv L, Lim P, Nhem S, Sem R, Khim N, Doung S, Mercereau-Puijalon O, Fandeur T: Large-scale malaria survey in Cambodia: novel insights on species distribution and risk factors. Malar J. 2007, 6: 37-10.1186/1475-2875-6-37.PubMedCentralCrossRefPubMed
11.
go back to reference Steenkeste N, Incardona S, Chy S, Duval L, Ekala MT, Lim P, Hewitt S, Sochantha T, Socheat D, Rogier C, Mercereau-Puijalon O, Fandeur T, Ariey F: Towards high- throughput molecular detection of Plasmodium: new approaches and molecular markers. Malar J. 2009, 8: 86-10.1186/1475-2875-8-86.PubMedCentralCrossRefPubMed Steenkeste N, Incardona S, Chy S, Duval L, Ekala MT, Lim P, Hewitt S, Sochantha T, Socheat D, Rogier C, Mercereau-Puijalon O, Fandeur T, Ariey F: Towards high- throughput molecular detection of Plasmodium: new approaches and molecular markers. Malar J. 2009, 8: 86-10.1186/1475-2875-8-86.PubMedCentralCrossRefPubMed
12.
go back to reference Hoyer S, Nguon S, Kim S, Habib N, Khim N, Sum S, Christophel EM, Bjorge S, Thomson A, Kheng S, Chea N, Yok S, Top S, Ros S, Sophal U, Thompson MM, Mellor S, Ariey F, Witkowski B, Yeang C, Yeung S, Duong S, Newman RD, Menard D: Focused screening and treatment (FSAT): a PCR-based strategy to detect malaria parasite carriers and contain drug resistant P. falciparum, Pailin, Cambodia. PLoS One. 2012, 7: e45797-10.1371/journal.pone.0045797.PubMedCentralCrossRefPubMed Hoyer S, Nguon S, Kim S, Habib N, Khim N, Sum S, Christophel EM, Bjorge S, Thomson A, Kheng S, Chea N, Yok S, Top S, Ros S, Sophal U, Thompson MM, Mellor S, Ariey F, Witkowski B, Yeang C, Yeung S, Duong S, Newman RD, Menard D: Focused screening and treatment (FSAT): a PCR-based strategy to detect malaria parasite carriers and contain drug resistant P. falciparum, Pailin, Cambodia. PLoS One. 2012, 7: e45797-10.1371/journal.pone.0045797.PubMedCentralCrossRefPubMed
13.
go back to reference Okell LC, Ghani AC, Lyons E, Drakeley CJ: Submicroscopic infection in Plasmodium falciparum-endemic populations: a systematic review and meta-analysis. J Infect Dis. 2009, 200: 1509-1517. 10.1086/644781.CrossRefPubMed Okell LC, Ghani AC, Lyons E, Drakeley CJ: Submicroscopic infection in Plasmodium falciparum-endemic populations: a systematic review and meta-analysis. J Infect Dis. 2009, 200: 1509-1517. 10.1086/644781.CrossRefPubMed
14.
go back to reference MalERA Consultative Group on Monitoring Evaluation and Surveillance: A research agenda for malaria eradication: monitoring, evaluation, and surveillance. PLoS Med. 2011, 8: e1000400-CrossRef MalERA Consultative Group on Monitoring Evaluation and Surveillance: A research agenda for malaria eradication: monitoring, evaluation, and surveillance. PLoS Med. 2011, 8: e1000400-CrossRef
15.
go back to reference Leang R, Barrette A, Bouth DM, Menard D, Abdur R, Duong S, Ringwald P: Efficacy of dihydroartemisinin-piperaquine for treatment of uncomplicated Plasmodium falciparum and Plasmodium vivax in Cambodia, 2008 to 2010. Antimicrob Agents Chemother. 2013, 57: 818-826. 10.1128/AAC.00686-12.PubMedCentralCrossRefPubMed Leang R, Barrette A, Bouth DM, Menard D, Abdur R, Duong S, Ringwald P: Efficacy of dihydroartemisinin-piperaquine for treatment of uncomplicated Plasmodium falciparum and Plasmodium vivax in Cambodia, 2008 to 2010. Antimicrob Agents Chemother. 2013, 57: 818-826. 10.1128/AAC.00686-12.PubMedCentralCrossRefPubMed
16.
go back to reference WHO: Malaria elimination. A field manual for low and moderate endemic countries. 2007, Geneva: World Health Organization WHO: Malaria elimination. A field manual for low and moderate endemic countries. 2007, Geneva: World Health Organization
17.
go back to reference Rakotonirina H, Barnadas C, Raherijafy R, Andrianantenaina H, Ratsimbasoa A, Randrianasolo L, Jahevitra M, Andriantsoanirina V, Menard D: Accuracy and reliability of malaria diagnostic techniques for guiding febrile outpatient treatment in malaria-endemic countries. Am J Trop Med Hyg. 2008, 78: 217-221.PubMed Rakotonirina H, Barnadas C, Raherijafy R, Andrianantenaina H, Ratsimbasoa A, Randrianasolo L, Jahevitra M, Andriantsoanirina V, Menard D: Accuracy and reliability of malaria diagnostic techniques for guiding febrile outpatient treatment in malaria-endemic countries. Am J Trop Med Hyg. 2008, 78: 217-221.PubMed
18.
go back to reference Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH: A review of malaria diagnostic tools: microscopy and rapid diagnostic test (RDT). Am J Trop Med Hyg. 2007, 77: 119-127.PubMed Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH: A review of malaria diagnostic tools: microscopy and rapid diagnostic test (RDT). Am J Trop Med Hyg. 2007, 77: 119-127.PubMed
19.
go back to reference Snounou G, Viriyakosol S, Zhu XP, Jarra W, Pinheiro L, Do Rosario VE, Thaithong S, Brown KN: High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction. Mol Biochem Parasitol. 1993, 61: 315-320. 10.1016/0166-6851(93)90077-B.CrossRefPubMed Snounou G, Viriyakosol S, Zhu XP, Jarra W, Pinheiro L, Do Rosario VE, Thaithong S, Brown KN: High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction. Mol Biochem Parasitol. 1993, 61: 315-320. 10.1016/0166-6851(93)90077-B.CrossRefPubMed
20.
go back to reference Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA: A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg. 1999, 60: 687-692.PubMed Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA: A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg. 1999, 60: 687-692.PubMed
21.
go back to reference Proux S, Suwanarusk R, Barends M, Zwang J, Price RN, Leimanis M, Kiricharoen L, Laochan N, Russell B, Nosten F, Snounou G: Considerations on the use of nucleic acid-based amplification for malaria parasite detection. Malar J. 2011, 10: 323-10.1186/1475-2875-10-323.PubMedCentralCrossRefPubMed Proux S, Suwanarusk R, Barends M, Zwang J, Price RN, Leimanis M, Kiricharoen L, Laochan N, Russell B, Nosten F, Snounou G: Considerations on the use of nucleic acid-based amplification for malaria parasite detection. Malar J. 2011, 10: 323-10.1186/1475-2875-10-323.PubMedCentralCrossRefPubMed
22.
go back to reference Maurin M: Real-time PCR as a diagnostic tool for bacterial diseases. Expert Rev Mol Diagn. 2012, 12: 731-754. 10.1586/erm.12.53.CrossRefPubMed Maurin M: Real-time PCR as a diagnostic tool for bacterial diseases. Expert Rev Mol Diagn. 2012, 12: 731-754. 10.1586/erm.12.53.CrossRefPubMed
23.
go back to reference Waters AP, McCutchan TF: Rapid, sensitive diagnosis of malaria based on ribosomal RNA. Lancet. 1989, 1: 1343-1346.CrossRefPubMed Waters AP, McCutchan TF: Rapid, sensitive diagnosis of malaria based on ribosomal RNA. Lancet. 1989, 1: 1343-1346.CrossRefPubMed
24.
go back to reference Snounou G, Singh B: Nested PCR analysis of Plasmodium parasites. Methods Mol Med. 2002, 72: 189-203.PubMed Snounou G, Singh B: Nested PCR analysis of Plasmodium parasites. Methods Mol Med. 2002, 72: 189-203.PubMed
25.
go back to reference Chou M, Kim S, Khim N, Chy S, Sum S, Dourng D, Canier L, Nguon C, Menard D: Performance of “VIKIA Malaria Ag Pf/Pan” (IMACCESS(R)), a new malaria rapid diagnostic test for detection of symptomatic malaria infections. Malar J. 2012, 11: 295-10.1186/1475-2875-11-295.PubMedCentralCrossRefPubMed Chou M, Kim S, Khim N, Chy S, Sum S, Dourng D, Canier L, Nguon C, Menard D: Performance of “VIKIA Malaria Ag Pf/Pan” (IMACCESS(R)), a new malaria rapid diagnostic test for detection of symptomatic malaria infections. Malar J. 2012, 11: 295-10.1186/1475-2875-11-295.PubMedCentralCrossRefPubMed
26.
go back to reference Farrugia C, Cabaret O, Botterel F, Bories C, Foulet F, Costa JM, Bretagne S: Cytochrome b gene quantitative PCR for diagnosing Plasmodium falciparum infection in travelers. J Clin Microbiol. 2011, 49: 2191-2195. 10.1128/JCM.02156-10.PubMedCentralCrossRefPubMed Farrugia C, Cabaret O, Botterel F, Bories C, Foulet F, Costa JM, Bretagne S: Cytochrome b gene quantitative PCR for diagnosing Plasmodium falciparum infection in travelers. J Clin Microbiol. 2011, 49: 2191-2195. 10.1128/JCM.02156-10.PubMedCentralCrossRefPubMed
27.
go back to reference Haanshuus CG, Mohn SC, Morch K, Langeland N, Blomberg B, Hanevik K: A novel, single-amplification PCR targeting mitochondrial genome highly sensitive and specific in diagnosing malaria among returned travellers in Bergen Norway. Malar J. 2012, 12: 26-CrossRef Haanshuus CG, Mohn SC, Morch K, Langeland N, Blomberg B, Hanevik K: A novel, single-amplification PCR targeting mitochondrial genome highly sensitive and specific in diagnosing malaria among returned travellers in Bergen Norway. Malar J. 2012, 12: 26-CrossRef
Metadata
Title
An innovative tool for moving malaria PCR detection of parasite reservoir into the field
Authors
Lydie Canier
Nimol Khim
Saorin Kim
Vincent Sluydts
Somony Heng
Dany Dourng
Rotha Eam
Sophy Chy
Chanra Khean
Kaknika Loch
Malen Ken
Hokkean Lim
Sovannaroath Siv
Sochantha Tho
Pascal Masse-Navette
Charlotte Gryseels
Sambunny Uk
Karel Van Roey
Koen Peeters Grietens
Mao Sokny
Boukheng Thavrin
Char Meng Chuor
Vincent Deubel
Lies Durnez
Marc Coosemans
Didier Ménard
Publication date
01-12-2013
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2013
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/1475-2875-12-405

Other articles of this Issue 1/2013

Malaria Journal 1/2013 Go to the issue
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.