EVALUATION OF TWO NESTED PCR-BASED DIAGNOSTIC ASSAYS FOR PLASMODIUM FALCIPARUM INFECTION
Keywords:Nested polymerase chain reaction, Plasmodium falciparum, Malaria, Thailand
Â Objective: The majority of malaria cases and deaths are caused by Plasmodium falciparum. The rapid and accurate diagnosis is very important for malaria treatment and control. The aim of this study was to evaluate two nested polymerase chain reaction (PCR)-based methods (protocol A and protocol B) for P. falciparum infection, diagnosis in Thailand.
Methods: A total of 90 dried blood spot samples were investigated. The samples composed of P. falciparum-, Plasmodium vivax-infected blood and normal human blood samples. The microscopic examination was used as gold standard.
Results: The results showed the sensitivity of 100/83.33%, specificity of 100/100%, and accuracy of 100/94.44% for protocol A and protocol B, respectively. The analytical sensitivity of protocol A and protocol B was 0.625 and 6.25 parasites/Î¼l, respectively. The comparison among microscopic examination, protocol A and protocol B by statistical analysis, found that they were not a significant difference. The agreements between each method were good. The kappa value between protocol A and protocol B was 0.87, protocol A and microscopy was 1.00, and protocol B and microscopy was 0.87.
Conclusion: The results demonstrated that protocol A should be used for further development of P. falciparum diagnosis in Thailand, especially in case of low parasitemia such as asymptomatic infection and for screening blood donors.
WHO. World Malaria Report 2016. Geneva: World Health Organization; 2016.
Hora R, Kapoor P, Thind KK, Mishra PC. Cerebral malaria-clinical manifestations and pathogenesis. Metab Brain Dis 2016;31:225-37.
Mungthin M, Suwandittakul N, Chaijaroenkul W, Rungsrihirunrat K, Harnyuttanakorn P, Seugorn A, et al. The patterns of mutation and amplification of Plasmodium falciparum pfcrt and pfmdr1 genes in Thailand during the year 1988 to 2003. Parasitol Res 2010;107(3):539-45.
Na-Bangchang K, Muhamad P, Ruaengweerayut R, Chaijaroenkul W, Karbwang J. Identification of resistance of Plasmodium falciparum to artesunate-mefloquine combination in an area along the Thai-Myanmar border: Integration of clinico-parasitological response, systemic drug exposure, and in vitro parasite sensitivity. Malar J 2013;12(1):263.
Ndao M, Bandyayera E, Kokoskin E, Gyorkos TW, MacLean JD, Ward BJ. Comparison of blood smear, antigen detection, and nested-PCR methods for screening refugees from regions where malaria is endemic after a malaria outbreak in Quebec, Canada. J Clin Microbiol 2004;42(6):2694-700.
Ayoola A, Adewumi S, Oyedele O, Saidu B, Mustapha O, Adetoro B, et al. Relevance of rapid immunodiagnostic test kits in malaria diagnosis. Int J Curr Pharm Res 2015;7(4):9-10.
Pakalapati D, Garg S, Middha S, Kochar A, Subudhi AK, Arunachalam BP, et al. Comparative evaluation of microscopy, OptiMAL(Â®) and 18S rRNA gene based multiplex PCR for detection of Plasmodium falciparum and Plasmodium vivax from field isolates of Bikaner, India. Asian Pac J Trop Med 2013;6(6):346-51.
Sharma J. Prevalence of malaria cases in tea garden areas of lakhimpur district, Assam. Int J Pharm Pharm Sci 2014;6(8):571-3.
Laban NM, Kobayashi T, Hamapumbu H, Sullivan D, Mharakurwa S, Thuma PE, et al. Comparison of a PfHRP2-based rapid diagnostic test and PCR for malaria in a low prevalence setting in rural southern Zambia: Implications for elimination. Malar J 2015;14(1):25.
Cunha MG, Medina TS, Oliveira SG, Marinho AN, PÃ³voa MM, Ribeiro-dos-Santos AK. Development of a polymerase chain reaction (PCR) method based on amplification of mitochondrial DNA to detect Plasmodium falciparum and Plasmodium vivax. Acta Trop 2009;111(1):35-8.
Stanis CS, Song BK, Chua TH, Lau YL, Jelip J. Evaluation of new multiplex PCR primers for the identification of Plasmodium species found in Sabah, Malaysia. Turk J Med Sci 2016;46(1):207-18.
Kongkasuriyachai D, Yongkiettrakul S, Kiatpathomchai W, Arunrut N. Loop-mediated isothermal amplification and LFD combination for detection of Plasmodium falciparum and Plasmodium vivax. Methods Mol Biol 2017;1572:431-43.
Snounou G, Viriyakosol S, Zhu XP, Jarra W, Pinheiro L, do Rosario VE, et al. High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction. Mol Biochem Parasitol 1993;61(2):315-20.
Anthony C, Mahmud R, Lau YL, Syedomar SF, Sri La Sri Ponnampalavanar S. Comparison of two nested PCR methods for the detection of human malaria. Trop Biomed 2013;30(3):459-66.
Li P, Zhao Z, Wang Y, Xing H, Parker DM, Yang Z, et al. Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys. Malar J 2014;13:175.
Yentur Doni N, Yildiz Zeyrek F, Seyrek A. Detection of Plasmodium using filter paper and nested PCR for patients with malaria in Sanliurfa, in Turkey. Malar J 2016;15(1):299.
Kritsiriwuthinan K, Ngrenngarmlert W. Molecular screening of Plasmodium infections among migrant workers in Thailand. J Vector Borne Dis 2011;48(4):214-8.
Taylor BJ, Martin KA, Arango E, Agudelo OM, Maestre A, Yanow SK. Real-time PCR detection of Plasmodium directly from whole blood and filter paper samples. Malar J 2011;10:244.
Imwong M, Nguyen TN, Tripura R, Peto TJ, Lee SJ, Lwin KM, et al. The epidemiology of subclinical malaria infections in South-East Asia: Findings from cross-sectional surveys in Thailand-Myanmar border areas, Cambodia, and Vietnam. Malar J 2015;14(1):385.
Arango EM, Samuel R, Agudelo OM, Carmona-Fonseca J, Maestre A, Yanow SK. Molecular detection of malaria at delivery reveals a high frequency of submicroscopic infections and associated placental damage in pregnant women from northwest Colombia. Am J Trop Med Hyg 2013;89(1):178-83.
Mahajan B, Zheng H, Pham PT, Sedegah MY, Majam VF, Akolkar N, et al. Polymerase chain reaction-based tests for pan-species and species-specific detection of human Plasmodium parasites. Transfusion 2012;52(9):1949-56.
Ebrahimzadeh A, Fouladi B, Fazaeli A. High rate of detection of mixed infections of Plasmodium vivax and Plasmodium falciparum in South-East of Iran, using nested PCR. Parasitol Int 2007;56(1):61-4.
Mehlotra RK, Lorry K, Kastens W, Miller SM, Alpers MP, Bockarie M, et al. Random distribution of mixed species malaria infections in Papua New Guinea. Am J Trop Med Hyg 2000;62(2):225-31.
Rubio JM, Benito A, Berzosa PJ, Roche J, Puente S, Subirats M, et al. Usefulness of seminested multiplex PCR in surveillance of imported malaria in Spain. J Clin Microbiol 1999;37(10):3260-4.
Hsiang MS, Lin M, Dokomajilar C, Kemere J, Pilcher CD, Dorsey G, et al. PCR-based pooling of dried blood spots for detection of malaria parasites: Optimization and application to a cohort of Ugandan children. J Clin Microbiol 2010;48(10):3539-43.
Coleman RE, Sattabongkot J, Promstaporm S, Maneechai N, Tippayachai B, Kengluecha A, et al. Comparison of PCR and microscopy for the detection of asymptomatic malaria in a Plasmodium falciparum/Vivax endemic area in Thailand. Malar J 2006;5:121.
RÃ¥dstrÃ¶m P, Knutsson R, Wolffs P, LÃ¶venklev M, LÃ¶fstrÃ¶m C. Pre-PCR processing: Strategies to generate PCR-compatible samples. Mol Biotechnol 2004;26(2):133-46.
Steenkeste N, Incardona S, Chy S, Duval L, Ekala MT, Lim P, et al. Towards high-throughput molecular detection of Plasmodium: New approaches and molecular markers. Malar J 2009;8(1):86.
Gupta B, Gupta P, Sharma A, Singh V, Dash AP, Das A. High proportion of mixed-species Plasmodium infections in India revealed by PCR diagnostic assay. Trop Med Int Health 2010;15(7):819-24.
Bin Dajem SM. Molecular investigation of mixed malaria infections in southwest Saudi Arabia. Saudi Med J 2015;36(2):248-51.
How to Cite
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.