Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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Comparative Assessment of Diagnostic Performance of Cytochrome Oxidase Multiplex PCR and 18S rRNA Nested PCR

  • Received : 2022.01.02
  • Accepted : 2022.05.16
  • Published : 2022.08.31
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Abstract

Malaria elimination and control require prompt and accurate diagnosis for treatment plan. Since microscopy and rapid diagnostic test (RDT) are not sensitive particularly for diagnosing low parasitemia, highly sensitive diagnostic tools are required for accurate treatment. Molecular diagnosis of malaria is commonly carried out by nested polymerase chain reaction (PCR) targeting 18S rRNA gene, while this technique involves long turnaround time and multiple steps leading to false positive results. To overcome these drawbacks, we compared highly sensitive cytochrome oxidase gene-based single-step multiplex reaction with 18S rRNA nested PCR. Cytochrome oxidase (cox) genes of P. falciparum (cox-III) and P. vivax (cox-I) were compared with 18S rRNA gene nested PCR and microscopy. Cox gene multiplex PCR was found to be highly specific and sensitive, enhancing the detection limit of mixed infections. Cox gene multiplex PCR showed a sensitivity of 100% and a specificity of 97%. This approach can be used as an alternative diagnostic method as it offers higher diagnostic performance and is amenable to high throughput scaling up for a larger sample size at low cost.

Keywords

Acknowledgement

Authors are thankful to the Director, ICMR-National Institute of Malaria Research, Delhi, India, for providing laboratory facilities.

References

  1. World Health Organization. World Malaria Report 2020: 20 Years of Global Progress and Challenges. Geneva, Switzerland. World Health Organization. 2020. https://apps.who.int/iris/handle/10665/337660
  2. Tangpukdee N, Duangdee C, Wilairatana P, Krudsood S. Malaria diagnosis: a brief review. Korean J Parasitol 2009; 47: 93-102. https://doi.org/10.3347/kjp.2009.47.2.93
  3. Kumar N, Singh JP, Pande V, Mishra N, Srivastava B, Kapoor R, Valecha N, Anvikar AR. Genetic variation in histidine rich proteins among Indian Plasmodium falciparum population: possible cause of variable sensitivity of malaria rapid diagnostic tests. Malaria J 2012; 11: 1-7. https://doi.org/10.1186/1475-2875-11-298
  4. Ochola LB, Vounatsou P, Smith T, Mabaso ML, Newton CR. The reliability of diagnostic techniques in the diagnosis and management of malaria in the absence of a gold standard. Lancet Infect Dis 2006; 6: 582-588. https://doi.org/10.1016/S1473-3099(06)70579-5
  5. Das A, Anvikar AR, Cator LJ, Dhiman RC, Eapen A, Mishra N, Nagpal BN, Nanda N, Raghavendra K, Read AF, Sharma SK, Singh OP, Singh V, Sinnis P, Srivastava HC, Sullivan SA, Sutton PL, Thomas MB, Carlton JM, Valecha N. Malaria in India: the center for the study of complex malaria in India. Acta Trop 2012; 121: 267-273. https://doi.org/10.1016/j.actatropica.2011.11.008
  6. Putaporntip C, Buppan P, Jongwutiwes S. Improved performance with saliva and urine as alternative DNA sources for malaria diagnosis by mitochondrial DNA-based PCR assays. Clin Microbiol Infect 2011; 17: 1484-1491. https://doi.org/10.1111/j.1469-0691.2011.03507.x
  7. Kumar N, Pande V, Bhatt R, Shah NK, Mishra N, Srivastava B, Valecha N, Anvikar AR. Genetic deletion of HRP2 and HRP3 in Indian Plasmodium falciparum population and false negative malaria rapid diagnostic test. Acta Trop 2013; 125: 119-121. https://doi.org/10.1016/j.actatropica.2012.09.015
  8. Mangold KA, Manson RU, Koay ES, Stephens L, Regner M, Thomson RB, Peterson LR, Kaul KL. Real-time PCR for detection and identification of Plasmodium spp. J Clin Microbiol 2005; 43: 2435-2440. https://doi.org/10.1128/JCM.43.5.2435-2440.2005
  9. Ocker R, Prompunjai Y, Chutipongvivate S, Karanis P. Malaria diagnosis by loop-mediated isothermal amplification (LAMP) in Thailand. Rev Inst Med Trop Sao Paulo 2016; 58: 27. https://doi.org/10.1590/S1678-9946201658027
  10. Yatsushiro S, Yamamura S, Yamaguchi Y, Shinohara Y, Tamiya E, Horii T, Baba Y, Kataoka M. Rapid and highly sensitive detection of malaria-infected erythrocytes using a cell microarray chip. PLoS One 2010, 5: e13179. https://doi.org/10.1371/journal.pone.0013179
  11. Cunha MG, Medina TS, Oliveira SG, Marinho AN, Povoa 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: 35-38. https://doi.org/10.1016/j.actatropica.2009.02.003
  12. Snounou G, Viriyakosol S, Jarra W, Thaithong S, Brown KN. Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections. Mol Biochem Parasitol 1993; 58: 283-292. https://doi.org/10.1016/0166-6851(93)90050-8
  13. Kumari P, Sinha S, Gahtori R, Yadav CP, Pradhan MM, Rahi M, Pande V, Anvikar AR. Prevalence of asymptomatic malaria parasitemia in Odisha, India: a challenge to malaria elimination. Am J Trop Med Hyg 2020; 103:1510-1516. https://doi.org/10.4269/ajtmh.20-0018
  14. Mohammed H, Kassa M, Mekete K, Assefa A, Taye G, Commons RJ. Genetic diversity of the msp-1, msp-2, and glurp genes of Plasmodium falciparum isolates in Northwest Ethiopia. Malaria J 2018; 17: 1-8. https://doi.org/10.1186/s12936-018-2540-x
  15. Imwong M, Pukrittayakamee S, Gruner AC, Renia L, Letourneur F, Looareesuwan S, White NJ, Snounou G. Practical PCR genotyping protocols for Plasmodium vivax using Pvcs and Pvmsp1. Malaria J 2005; 4: 1-13. https://doi.org/10.1186/1475-2875-4-20
  16. Hanscheid T, Grobusch MP. How useful is PCR in the diagnosis of malaria? Trends Parasitol 2002; 18: 395-398. https://doi.org/10.1016/s1471-4922(02)02348-6
  17. Hanscheid T, Valadas E. Poor accuracy of rapid diagnostic tests and misdiagnosis of imported malaria: are PCR-based reference laboratories the answer? J Clin Microbiol 2002; 40: 736-737. https://doi.org/10.1128/JCM.40.2.736-737.2002
  18. 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. Malaria J 2013; 12: 1-8. https://doi.org/10.1186/1475-2875-12-26
  19. Preiser PR, Wilson RJ, Moore PW, McCready S, Hajibagheri MA, Blight KJ, Strath M, Williamson DH. Recombination associated with replication of malarial mitochondrial DNA. EMBO J 1996; 15: 684-693. https://doi.org/10.1002/j.1460-2075.1996.tb00401.x
  20. Polley SD, Mori Y, Watson J, Perkins MD, Gonzalez IJ, Notomi T, Chiodini PL, Sutherland CJ. Mitochondrial DNA targets increase sensitivity of malaria detection using loop-mediated isothermal amplification. J Clin Microbiol 2010; 48: 2866-2871. https://doi.org/10.1128/JCM.00355-10