A Triplex Nested Polymerase Chain Reaction for Simultaneously Diagnosis of Plasmodium, Babesia, and Toxoplasma in Blood to Promote Safety of Blood Transfusion

Document Type : Original Article (s)

Authors

1 Associate Professor, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 MSc Student, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences AND Isfahan Blood Transfusion Organization, Isfahan, Iran

Abstract

Background: Diagnosis and remove of blood transmittable infections has a vital importance in blood transfusion. The aim of this study was to concurrently assess the presence of protozoan parasites plasmodium, toxoplasma, and babesia in samples of blood transfusion.Methods: In this experimental study, the development of a triplex nested-Polymerase chain reaction (PCR) for extol of blood transfusion process at 2020 in Isfahan City, Iran, was done. After selecting the genus specific primers, at the first step, nested PCRs were set up for each parasite specifically at the genus level using blood samples with a determined number of each parasite, and then multiplex nested PCR was set up.Findings: The nested PCR triple reaction (TN-PCR) has a higher diagnostic validity compared to the microscopic and enzyme-linked immunosorbent assay (ELISA) methods. Sensitivity, specificity, and positive and negative predictive values were as 100%, 100%, 100%, and 100%, respectively, for detection Plasmodium, 100%, 91%, 92%, and 100%, for Toxoplasma, and 94%, 100%, 100%, and 89% for Babesia. The validity of this test for Plasmodium was the highest compared to the other two parasites.Conclusion: Based on the results of this study, the TN-PCR method can simultaneously identify one, two or all three dangerous pathogens in biological samples as well as blood samples in the blood transfusion process.

Keywords

References

  1. Keng Ho P, Loke J. Clinical blood transfusion. Singapore: Ministry of Health; 2011. p. 23.
  2. Karimi G, Mardani A, Zadsar M. Prevalence of Toxoplasma gondii among Iranian blood donors: A narrative review article. Iran J Parasitol 2016; 11(1): 10-8.
  3. Karimi G, Gharehbaghian A, Tafti MF, Vafaiyan V. Emerging infectious threats to the blood supply: Seroepidemiological studies in iran - a review. Transfus Med Hemother 2013; 40(3): 210-7.
  4. Sputin A. Medical Parasitology for medical students: 1st ed. Tehran, Iran: Khosravi Publications; 2010. [In Persian].
  5. World Health Organization. Screening donated blood for transfusion-transmissible infections: Recommendations. Geneva, Switzerland: World Health Organization; 2010.
  6. Sundar P, Mahadevan A, Jayshree RS, Subbakrishna DK, Shankar SK. Toxoplasma seroprevalence in healthy voluntary blood donors from urban Karnataka. Indian J Med Res 2007; 126(1): 50-5.
  7. Seed CR, Kitchen A, Davis TM. The current status and potential role of laboratory testing to prevent transfusion-transmitted malaria. Transfus Med Rev 2005; 19(3): 229-40.
  8. Singh G, Sehgal R. Transfusion-transmitted parasitic infections. Asian J Transfus Sci 2010; 4(2): 73-7.
  9. Lima GFMC, Arroyo Sanchez MC, Levi JE, Fujimori M, Da Cruz CL, Sanchez AR, et al. Asymptomatic infections in blood donors harbouring Plasmodium: an invisible risk detected by molecular and serological tools. Blood Transfus 2018; 16(1): 17-25.
  10. Verra F, Angheben A, Martello E, Giorli G, Perandin F, Bisoffi Z. A systematic review of transfusion-transmitted malaria in non-endemic areas. Malar J 2018; 17(1): 36.
  11. Hill DE, Dubey JP. Toxoplasma gondii as a Parasite in Food: Analysis and Control. Microbiol Spectr 2016; 4(4).
  12. Ghadamgahi F, Bahadoran M, Shariat-Bahadori E, Ahmadi-Ahvaz N, Ghadrdoost B, Hejazi SH. Study of serological toxoplasmosis and risk factors associated with infection in women referred to labs of northern Tehran, Iran. J Isfahan Med Sch 2013; 31(248): 1257-66. [In Persian].
  13. Siegel SE, Lunde MN, Gelderman AH, Halterman RH, Brown JA, Levine AS, et al. Transmission of toxoplasmosis by leukocyte transfusion. Blood 1971; 37(4): 388-94.
  14. Alishahi A, Fallahian M, Fallahian F. Medical Parasitology. 1st ed. Tehran, Iran: Jameah Negar Publications; 2014. [In Persian].
  15. Tonnetti L, Townsend RL, Deisting BM, Haynes JM, Dodd RY, Stramer SL. The impact of Babesia microti blood donation screening. Transfusion 2019; 59(2): 593-600.
  16. Leiby DA. Babesiosis and blood transfusion: Flying under the radar. Vox Sang 2006; 90(3): 157-65.
  17. Mukry SN, Saud M, Sufaida G, Shaikh K, Naz A, Shamsi TS. laboratory diagnosis of malaria: Comparison of manual and automated diagnostic tests. Can J Infect Dis Med Microbiol 2017; 2017: 9286392.
  18. Ghasemi FS, Hooshyar H. Laboratory diagnosis of Toxoplasmosis and its importance. J Med Counc I R Iran 2014; 32(1): 65-77. [In Persian].
  19. Nahravanian H, Eskandarian AA. Malaria and malaria parasites. 1st ed. Tehran, Iran: Khosravi Publications; 2005. [In Persian].
  20. Karami M, Mahbod SAA, Shad Del M. Comparison between IFA and ELISA technique with PCR method in the diagnosis of Toxoplasma gondi parasite in infants admitted at taleghani hospital. Annals of Military and Health Sciences Research 2007; 5(1): 1141-9. [In Persian].
  21. Yentur DN, Yildiz ZF, 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.
  22. Mousavi M, Saravani R, Jafari MM, Shahrakipour M, Sekandarpour S. Detection of Toxoplasma gondii in diabetic patients using the nested PCR assay via RE and B1 genes. Jundishapur J Microbiol 2016; 9(2): e29493.
  23. Kim JY, Cho SH, Joo HN, Tsuji M, Cho SR, Park IJ, et al. First case of human babesiosis in Korea: Detection and characterization of a novel type of Babesia sp. (KO1) similar to ovine babesia. J Clin Microbiol 2007; 45(6): 2084-7.
  24. Zakeri S, Najafabadi ST, Zare A, Djadid ND. Detection of malaria parasites by nested PCR in south-eastern, Iran: Evidence of highly mixed infections in Chahbahar district. Malar J 2002; 1: 2.
  25. Lam WY, Yeung AC, Tang JW, Ip M, Chan EW, Hui M, et al. Rapid multiplex nested PCR for detection of respiratory viruses. J Clin Microbiol 2007; 45(11): 3631-40.
  26. Poschl B, Waneesorn J, Thekisoe O, Chutipongvivate S, Karanis P. Comparative diagnosis of malaria infections by microscopy, nested PCR, and LAMP in northern Thailand. Am J Trop Med Hyg 2010; 83(1): 56-60.
  27. Arifin S, Fitri LE, Sujuti H, Hermansyah B, Endharti AT, Burhan N, et al. Sensitivity and specificity of nested PCR for diagnosing malaria: Cases in several areas of Indonesia. J Trop Life Sci 2018; 8(21): 172-6.
  28. Saki J, Foroutan M, Khodkar I, Khodadadi A, Nazari L. Seroprevalence and molecular detection of Toxoplasma gondii in healthy blood donors in southwest Iran. Transfus Apher Sci 2019; 58(1): 79-82.

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History

  • Receive Date: 08 December 2020
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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