Evaluation of the detection of Plasmodium falciparum infection in Urine, Plasma, Serum, and Dried Blood Spots using real-time Polymerase Chain Reaction among Febrile patients in Lagos, Nigeria
https://dx.doi.org/10.4314/njpar.v44i1.7
Keywords:
diagnostic performances, Microscopy, Real-time PCR, Plasmodium speciesAbstract
Access to timely and accurate diagnostic tests has a significant impact on the management of malaria disease which is a global concern. Polymerase chain reaction (PCR) detection of Plasmodium DNA is highly sensitive in diagnosing malaria. The specimen of choice for this assay has been whole blood samples from patients with malaria caused by Plasmodium species. Nucleic acids can also be detected in urine, serum, plasma, and Dried Blood Spots (DBS) samples but there are few studies describing the diagnostic performance of PCR. Therefore, this study was aimed at evaluating the performance of realtime PCR (qPCR) in detecting malaria parasite DNA in serum, plasma, urine, and DBS. A cross-sectional study was conducted among 146 patients that attended the clinic at Bayeku, Oreta, Imota, Ijede, Agura Primary Health Centres (PHC) and Ikorodu General hospital of Lagos State. Urine samples and a total of 5 ml of blood were collected from each participant and made into dried blot spots, plasma, and serum. The samples were screened and assayed for Plasmodium falciparum by microscopy and Multiplex qPCR respectively. The sensitivity of qPCR using plasma, serum and urine specimens were 100%, 87%, and 52.6% respectively, while the specificity was 82%, 87.5% and 80% respectively. Parasite detection by microscopy showed greater agreement with detection by qPCR in serum (79.4%) than qPCR from plasma (75%) or urine (58.3%). In conclusion, malaria detection using qPCR assay on plasma has high sensitivity and can be used as an alternative to microscopy.
References
World Health Organization (2012). Management of Severe Malaria, Global Malaria Programme, a Practical Handbook. 3rd ed. Geneva: World Health Organization
White, N. J. (2004). Antimalarial drug resistance. The Journal of clinical investigation 113(8):1084-1092.
Tangpukdee, N., Duangdee, C., Wilairatana, P. and Krudsood S. (2009). Malaria diagnosis: a brief review. Korean Journal of Parasitology 47(2):93-102.
Moody, A. (2002) Rapid diagnostic tests for malaria parasites. Clinical Microbiology Review 15(1):66-78.
Bharti, A. R., Letendre, S. L., Patra, K. P., Vinetz, J. M. and Smith, D. M. (2009). Malaria diagnosis by a polymerase chain reaction-based assay using a pooling strategy. American Journal of Tropical Medical Hygiene 81(5):754-757.
Fançony, C., Sebastião, Y. V., Pires, J. E., Gamboa, D. and Nery, S. V. (2013). Performance of microscopy and RDTs in the context of a malaria prevalence survey in Angola: a comparison using PCR as the gold standard. Malaria Journal 12:284.
Murray, C. K., Gasser, R. A Jr., Magill, A. J., and Miller, R. S. (2008). Update on rapid diagnostic testing for malaria. Clinical Microbiology Review 21(1):97-110.
Perandin. F., Manca, N., Calderaro, A., Piccolo, G., Galati, L., Ricci, L., Medici, M. C., Arcangeletti, M. C., Snounou, G., Dettori, G. and Chezzi, C. (2004). Development of a real-time PCR assay for detection of Plasmodium falciparum, Plasmodium vivax, and Plasmodium ovale for routine clinical diagnosis. Journal of Clinical Microbiology 42(3):1214-1219.
Johnston, S. P., Pieniazek, N. J.,Xayavong, M. V., Slemenda, S. B., Wilkins, P. P. and da Silva A, J. (2006). PCR as a confirmatory technique for
laboratory diagnosis of malaria. Journal of Clinical Microbiology 44(3):1087-1089.
Davis, C, Nwakanma, N. G., Michael, W., Sarah, C., Filip, D., Elissa, M., Emily, L., David, J. (2009). Quantitative Detection of Plasmodium falciparum DNA in Saliva, Blood, and Urine. Journal of Infective Disease 199 (111):1567–1574
Buppan, P. , Putaporntip, C. , Pattanawong, U., Seethamchai, S. and Jongwutiwes, S. (2010). Comparative detection of Plasmodium vivax and Plasmodium falciparum DNA in saliva and urine samples from symptomatic malaria patients in a low endemic area. Malaria Journal 9: 72
Gatti, S. M., Gramegna, Z., Bisoffi, A., Raglio, M., Gulletta, C., Klersy, A., Bruno, R., Maserati, S. M. and Scaglia, A. (2007). comparison of three diagnostic techniques for malaria: a rapid diagnostic test (NOWMalaria), PCR and microscopy. Annals of Tropical Medical Parasitology
:195–204.
Ojurongbe, O., Adegbosin, O. O., Taiwo, S. S., Alli, O. A., Olowe, O.A., Ojurongbe, T. A., Bolaji, O. S. and Adeyeba, O. A. (2013). Assessment of Clinical Diagnosis, Microscopy, Rapid Diagnostic Tests, and Polymerase Chain Reaction in the Diagnosis of Plasmodium falciparum in Nigeria.
Malaria Research Treatment 2013:308069.
Azikiwe, C. C., Ifezulike, C. C., Siminialayi, I. M., Amazu, L. U., Enye, J. C. and Nwakwunite, O. E. (2012). A comparative laboratory diagnosis of
malaria: microscopy versus rapid diagnostic test kits. Asian Pacific Journal of Tropical Biomedical 2(4):307-10.
Mharakurwa, S., Simoloka, C., Thuma, P. E., Shiff, C. J., Sullivan, D. J. (2006). PCR detection of Plasmodium falciparum in human urine and saliva samples. Malaria Journal 5:103
Bharti, A. R., Patra, K. P., Chuquiyauri, R., Kosek, M., Gilman, R. H., LlanosCuentas, A., Vinetz, J. M. (2007). Polymerase chain reaction detection of Plasmodium vivax and Plasmodium falciparum DNA from stored serum samples: implications for retrospective diagnosis of malaria. The American Journal of Tropical Medical Hygiene 7(3):444-446.
World Health Organisation (2009). Guidelines for Using HIV Testing Technologies in Surveillance, Selection, Evaluation, and Implementation: Update, Geneva: World Health Organization.
Oluwagbemiga, O.,Aina, C. O.,Agomo,Y. A., Olukosi, H. I., Okoh, B. A., Iwalokun, K. N., Egbuna, A. B., Orok, O. A., Veronica, N. V., Enya, Samuel K. A., Margaret, O. A. and Philip, U. A. (2013). Malariometric Survey of Ibeshe Community in Ikorodu, Lagos State: Dry Season.
Malaria Research and Treatment 2013: 01- 07
Monica, C. (2009). District laboratory practice in tropical countries. Part one (2nd edition). Cambridge, UK: Cambridge University Press.
Waggoner, J. J., Abeynayake, J., Sahoo, M. K., Gresh, L., Tellez, Y., Gonzalez, K., Ballesteros, G., Balmaseda, A., Karunaratne, K., Harris, E. and Pinsky, B. A. (2013) Development of an internally controlled real-time reverse transcriptase PCR assay for pan-dengue virus detection and comparison of four molecular dengue virus detection assays. Journal of Clinical Microbiology 51:2172–2181
Waggoner, J. J., Balassian, I., Abeynayake, J., Sahoo, M. K., MohamedHadle,A., Liu, Y., Magalha, J. and Pinsky, B. A. (2014). Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of NucleicAcidAmplification Methods for the Diagnosis of Leptospirosis.
PLOS ONE 9(11): e112356
Waggoner, J. J, Okangba, C., MohamedHadley A., Lefterova, M. I., Banaei, N., Oyibo, W. and Pinsky, B. A. (2015). Molecular Testing for Plasmodium falciparum by Use of Serum or Plasma and Comparison with Microscopy and Rapid Diagnostic Testing in Febrile Nigerian
Patients. Journal of Clinical Microbiology 53(11):3596-3600.
Lamikanra, A. A., Dobano, C., Jimenez, A., Nhabomba, A., Tsang, H. P., Guinovart, C., Manaca, M. N., Quinto, L., Aguilar, R., Cistero, P., Alonso, P. L., Roberts, D. J. and Mayor, A. (2012). A direct comparison of real-time PCR on plasma and blood to detect Plasmodium falciparum infection in children. Malaria Journal 11:201.
Andrade, B. B., Reis-Filho, A., Barros, A. M., Souza-Neto S. M., Nogueira, L. L. and Fukutani, K. F. (2010). Towards a precise test for malaria diagnosis in the Brazilian Amazon: Comparison among field microscopy, a rapid diagnostic test, nested PCR, and a computational expert system based on artificial neural networks. Malaria Journal 9:117
Lucy, C. O., Azra, C., Ghani, E. L., and Chris J. D. (2009). Submicroscopic Infection in Plasmodium falciparumEndemic Populations: A Systematic Review and Meta-Analysis. Journal of Infective Disease 200 (10) 15 :1509–1517
Boonma, P., Christensen, P. R., Suwanarusk, R., Price, R. N., Russell, B., and Lek-Uthai, U. (2007). Comparison of
three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum. Malaria Journal
:124.
Singh, B. (1997). Molecular methods for diagnosis and epidemiological studies of parasitic infections. International Journal of
parasitology 27:1135-1145
Golassa, L., Enweji, N., Erko, B. and Aseffa A. (3013). Detection of a substantial number of submicroscopic Plasmodium falciparum infections by polymerase chain reaction:Apotential threat to malaria control and diagnosis in Ethiopia. Malaria Journal 12: 352
Steenkeste, N., Incardona, S., Chy. S., Duval, L., Ekala, M., Lim, P., Hewitt, S., Sochantha, T., Sochea, D., Rogier, C., Mercereau-Puijalon, O., Fandeur, T. and Ariey, F. (2009). Towards high-throughput molecular detection of Plasmodium: new approaches and molecular markers. Malaria Journal 8: 86
Pembele, G., Rivero, L. and Fraga, J. (2015). Detection and Species Identification of Malaria Parasites by Nested-PCR: Comparison with Light Microscopy and with SD BIOLINE Malaria Ag Test in Luanda, Angola. International Journal of Tropical Disease & Health 10(1): 1-13
Oyedeji, S. I., Odoh, I. M., Ojerinde, A. O. and Awobode, H. O. (2021). Investigation of the sequence profile of the Plasmodium falciparum 18SrRNA diagnostic target in isolates from naturally infected children with uncomplicated malaria. Nigeria Journal of Parasitology 42(2):242-250
Berzosa, P., De Lucio, A., Romay-Barja, M., Zaida Herrador, V. G., Luz, G., Amalia, F., Maria, S., Policarpo, N., Basilio, V., Matilde. R. and Agustín, B. (2018). Comparison of three diagnostic methods (microscopy, RDT, and PCR) for the detection of malaria parasites in representative samples from Equatorial Guinea. Malaria Journal 17(333): 01-12
Moura, S., Fançony, C., Mirante, C., Neves, M., Bernardino, L., Fortes, F., Sambo, M. R. and Brito, M. (2014). Impact of a training course on the quality of malaria diagnosis by microscopy inAngola. Malaria Journal 13(437):[Internet]
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Nigerian Journal of Parasitology
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
