Comparison of RT.PCR and ELISA Methods in the Diagnosis of Human Cytomegalovirus in Kidney Transplant Patients
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Abstract
Background: In kidney transplant recipients prone to infections like cytomegalovirus (CMV), a vital need for an accurate diagnostic method is evident. This study at Khorshid Laboratory in Tehran rigorously compares real-time PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) for CMV detection in transplant patients.
Methods: In January to March 1400, 70 kidney transplant recipients were assessed for CMV DNA using RT-PCR and concurrent ELISA tests, with statistical analysis aided by SPSS.
Results: In kidney transplant patients (average age: 49.40 ± 13.64 years), 4.3% tested positive for CMV via PCR. Strong correlation between serological and molecular methods. IgG and IgM antibody detection both showed high sensitivity and specificity, advocating for efficient CMV diagnosis at Khorshid Laboratory, Tehran.
Conclusion: This study emphasizes the need for a quick and efficient CMV diagnostic approach in kidney transplant patients. The strong correlation between molecular and serological methods favors using the faster RT-PCR method, crucial for timely management, especially with the increasing age-related CMV incidence. The findings strongly recommend RT-PCR integration for enhanced sensitivity in kidney transplant patients at Khorshid Laboratory, Tehran.
1. Kumar R, Ison MG. Opportunistic infections in transplant patients. Infect Dis Clin 2019; 33: 1143-57.
2. Ariza Heredia EJ, Nesher L, Chemaly RF. Cytomegalovirus diseases after hematopoietic stem cell transplantation: A mini-review. Cancer Lett 2014; 342:1-8.
3. Zhang S, Springer LE, Rao HZ, et al. Hematopoietic cell-mediated dissemination of murine cytomegalovirus is regulated by NK cells and immune evasion. PLoS Pathog 2021; 17.
4. Picone O, Vauloup‐Fellous C, Cordier AG, et al. A 2‐year study on cytomegalovirus infection during pregnancy in a French hospital. BJOG 2009; 116(6):818-23.
5. Hassan J, O’Neill D, Honari B, et al. Cytomegalovirus infection in ireland: seroprevalence, HLA Class I Alleles, and Implications. Medicine (Baltimore) 2016; 95(6):e2735.
6. Enders G, Daiminger A, Lindemann L, et al. Cytomegalovirus (CMV) seroprevalence in pregnant women, bone marrow donors and adolescents in Germany, 1996–2010. Med Microbiol Immunol 2012; 201(3):303-9.
7. Pembrey L, Raynor P, Griffiths P, et al. Seroprevalence of cytomegalovirus, Epstein Barr virus and varicella zoster virus among pregnant women in Bradford: a cohort study. PloSOne 2013; 8(11):e81881.
8. Abu-Madi MA, Behnke JM, Dabritz HA. Toxoplasma gondii seropositivity and coinfection with TORCH pathogens in highrisk patients from Qatar. Am J Trop Med Hyg 2010; 82(4):626-33.
9. Shaiegan M, Rasouli M, Zadsar M, et al. Meta-analysis of cytomegalovirus seroprevalence in volunteer blood donors and healthy subjects in Iran from 1992 to 2013. Iran J Basic Med Sci 2015; 18(7):627-34
10. Harari A, Zimmerli SC, Pantaleo G. Cytomegalovirus (CMV)-specific cellular immune responses. Hum Immunol 2004; 65: 500-6.
11. Rump K, Rahmel T, Rustige AM, et al. The aquaporin 3 promoter polymorphism− 1431 A/G is associated with acute graft rejection and cytomegalovirus infection in kidney recipients due to altered immune cell migration. Cells 2020; 9: 1421.
12. Shoup M, Ourahmane A, Ginsburg EP, et al. Substitution-inert polynuclear platinum compounds inhibit human cytomegalovirus attachment and entry. Antivir Res 2020; 184:104957
13. Liu Z, Yu Y, Liu H, et al. Establishment of a sandwich light-initiated chemiluminescence assay with double antigen for detecting human cytomegalovirus IgG antibody. J Med Virol 2021; 93:5025-32
14. Awadh N, Nyawale H, Chibwe E, et al. Serological markers of viral infections (rubella virus, human cytomegalovirus and arboviruses) among symptomatic pregnant women in rural and urban areas of Mwanza, Tanzania. Trop Med Infec Dis 2021; 6: 186-212.
15. Zavaglio F, Fiorina L, Suárez NM, et al. Detection of genotype-specific antibody responses to glycoproteins b and h in primary and non-primary human cytomegalovirus infections by peptide-based ELISA. Viruses 2021; 13:399-405.
16. Milavec M, Pavšiˇc J, Košir AB, et al. The performance of human cytomegalovirus digital PCR Reference Measurement Procedure in seven External Quality Assessment schemes over four years. Methods 2021; 25:107-12.
17. Lazar K, Rabe T, Goelz R, et al. Human cytomegalovirus reactivation during lactation: Impact of antibody kinetics and neutralization in blood and breast milk. Nutrients 2020; 12:338-41.
18. Hirsch HH, Lautenschlager I, Pinsky BA, et al. An international multicenter performance analysis of cytomegalovirus load tests. Clin Infect Dis 2013; 56(3):367-73.
19. Ziyaeyan M, Sabahi F. Human cytomegalovirus: infections and diagnosis. Iran J Med Sci 2008; 33(3):127-32.
20. Razonable RR, Humar A. Cytomegalovirus in solid organ transplant recipients- Guidelines of the american society of transplantation infectious diseases community of practice. Clin Transplant 2019; 33(9):e13512.
21. Eshaghi F. Bashi Zadeh Fakhar H, Ghane M, et al. Diagnostic evaluation of Streptococcus gallolyticus infection in patients with colon diseases by polymerase chain reaction (PCR) and culturing methods. Int J Cancer Manag 2020; 13(6):e101903.
22. Merchant EA, Flint K, Barouch DH, et al. Coinfection with coronavirus disease 2019, previously undiagnosed human immunodeficiency virus, Pneumocystis jirovecii pneumonia and cytomegalovirus pneumonitis, with possible immune reconstitution inflammatory syndrome. Idcases 2021; 24:e01153.
23. Covid19.who.int. 2021. WHO Coronavirus (COVID-19). [online].
24. Seyed-Alinaghi S, Afsahi AM, MohsseniPour M, et al. Late complications of COVID-19; a systematic review of current evidence. Arch Acad Emerg Med 2021; 9(1):e14.
25. Kotton NC, Kumar D, Caliendo AM, et al. Updated international consensus guidelines on the management of cytomegalovirus in solidorgan transplantation. Transplantation 2013; 96(4):333-60.
26. Mocarski ES, Shenk T, Pass RF. Cytomegalovirus. In: Knipe DM, Howley PM, ed, Fields Virology, Philadelphia, Lippicott Williams&Wilkins. 2007; PP:2701-2772.
27. Kouri V, Resik S, Enamorado A, et al. Longitudinal study of herpesviruses in kidney transplant recipients in Cuba. Clin Infect Dis 2003; 36(6):818-21.
28. Erice A. Resistance of human cytomegalovirus to antiviral drugs. Clin Microbiol Rev 1999; 12(2): 286-97.
29. Forman SJ, Zaia JA. Treatment and prevention of cytomegalovirus pneumonia after bone marrow transplantation: where do we stand? Blood 1994; 83:2392-8.
30. Enders G, Daiminger A, Lindemann L, et al. Cytomegalovirus (CMV) seroprevalence in pregnant women, bone marrowdonors and adolescents in Germany, 1996-2010. Med Microbiol Immunol 2012; 201(3):303-9
31. Pembrey L, Raynor P, Griffiths P, et al. Seroprevalence of cytomegalovirus, epstein barr virus and varicella zoster virus among pregnant women in bradford: a cohort study. PLoS One 2013; 8(11):e81881.
32. Abu-Madi MA, Behnke JM, Dabritz HA. Toxoplasma gondii seropositivity and co-infection with TORCH pathogens in high-risk patients from Qatar. Am J Trop Med Hyg 2010; 82(4):626-33.
33. Ali S, Khan FA, Mian AA, et al. Seroprevalence of cytomegalovirus, herpes simplex virus and rubella virus among pregnant women in KPK province of Pakistan. J Infect Dev Ctries 2014; 8(3):389-90.
34. Shaiegan M, Rasouli M, Zadsar M, et al. Meta‐analysis of cytomegalovirus sero-prevalence in volunteer blood donors and healthy subjects in Iran from 1992 to 2013. Iran J Basic Med Sci 2015; 18(7):627-34.
35. Motamedifar M, Hashemizadeh Z, Hadi N, et al. The study of seroprevalence of human cytomegalovirus in healthy blood donors referring to Fars blood bank. Hormozgan Med J 2009; 12(4):237-42.[In Persian].
36. Case IPTA. 237 coinfection with COVID-19 and cytomegalovirus in kidney transplant recipients–a case series. Transplant proceed 2021; 54(12):251-260.
37. Bal Z, Uyar M, Tutal E, et al. Cytomegalovirus Infectıon in renal transplant recipients: one center's experience. Transplant proceed 2013; 45(10):3520-3.
38. Chiasakul T, Townamchai N, Jutivorakool K, et al. Risk factors of cytomegalovirus disease in kidney transplant recipients: a single-center study in thailand. Transplant Proceed 2015; 47(8):2460-4.
39. Kalkan IH, Dağli U. What is the most accurate method for the diagnosis of cytomegalovirus (CMV) enteritis or colitis. Turk J Gastroenterol 2010; 21:83-6.
40. Jamalidoust M, Namayandeh M, Pouladfar G, et al. Post-Liver transplant cytomegalovirus (cmv) reactivation, graft, and patient survival rates in iranian population. Jundishapur J Microbiol 2021; 14(3):e112775.
41. Lakzayi E, Fozouni L, Pordeli H, et al. A Seromolecular Study on the prevalence of cytomegalovirus in pregnant women living in Golestan Province, Iran. Shiraz E-Med J 2020; 21(7):e96238.
42. Rahimzadeh G, Shahbaznejad L, Fahimzad A, et al. Seroepidemiology of cytomegalovirus in pregnant women in sari, Iran 2018-2020. J Mazandaran Univ Med Sci 2021; 31(202):149-154.
43. Fadhil HY. Correlation the cytomegalovirus (cmv) igm antibodies and viral dna presence with rheumatoid arthritis (ra) in iraqi patients. Iraqi J Sci 2019; 60(11):2383-2389.
44. Mohsin IH, Badday Hassan I, Daim Saleh MA. A comparative study of immunological and molecular techniques to diagnose human cytomegalovirus in renal failure patients in diyala governor. Ind J Forensic Med Toxicol 2022; 16(1):107-12.
45. Bahri T. Comparison of ELISA, antigenemia assay and nested pcr monitoring techniques for detection of cytomegalovirus infection. J Med Microb Diagn 2018, 7:1.
46. Miller H, Simpson P, Forman M, et al. A novel multiplexed enzyme-linked immunosorbent assay for the detection of igg seroreactivity to cytomegalovirus (CMV) UL144. J Clin Microbiol 2021; 59(8):e00964-21.
47. Shkhair H, Al-janabi O, Abed Kelkawi AH, et al. Comparative evaluation study of ELISA system and MINI-VIDAS system for detection of Cytomegalovirus IgM antibodies. J Pharm Sci Res 2018; 10(10):2549-2550.
48. Çolak D, Sa ̆glık ̇I, Mutlu D, et al. Comparison of two commercial quantitative cy-tomegalovirus (cmv) polymerase chain reaction tests calibrated by world health organization international cmv standard. Mikrobiyoloji Bul 2020; 54:257-265.
49. Stilp AC, König P, Scherer M, et al. Stable and inducible gene knockdown in primary human fibroblasts: a versatile tool to study the role of human cytomegalovirus host cell factors. In Human Cytomegaloviruses; Humana: New York,NY, USA.2021; pp. 115–132.
50. Aghajanian S, Rangani A, Alizadeh-Navaei R, et al. Epidemiology of cytomegalovirus among patients undergoing hemodialysis in Sari, Iran 2017-2018. J Mazandaran Univ Med Sci 2021, 31(199):179-185.
2. Ariza Heredia EJ, Nesher L, Chemaly RF. Cytomegalovirus diseases after hematopoietic stem cell transplantation: A mini-review. Cancer Lett 2014; 342:1-8.
3. Zhang S, Springer LE, Rao HZ, et al. Hematopoietic cell-mediated dissemination of murine cytomegalovirus is regulated by NK cells and immune evasion. PLoS Pathog 2021; 17.
4. Picone O, Vauloup‐Fellous C, Cordier AG, et al. A 2‐year study on cytomegalovirus infection during pregnancy in a French hospital. BJOG 2009; 116(6):818-23.
5. Hassan J, O’Neill D, Honari B, et al. Cytomegalovirus infection in ireland: seroprevalence, HLA Class I Alleles, and Implications. Medicine (Baltimore) 2016; 95(6):e2735.
6. Enders G, Daiminger A, Lindemann L, et al. Cytomegalovirus (CMV) seroprevalence in pregnant women, bone marrow donors and adolescents in Germany, 1996–2010. Med Microbiol Immunol 2012; 201(3):303-9.
7. Pembrey L, Raynor P, Griffiths P, et al. Seroprevalence of cytomegalovirus, Epstein Barr virus and varicella zoster virus among pregnant women in Bradford: a cohort study. PloSOne 2013; 8(11):e81881.
8. Abu-Madi MA, Behnke JM, Dabritz HA. Toxoplasma gondii seropositivity and coinfection with TORCH pathogens in highrisk patients from Qatar. Am J Trop Med Hyg 2010; 82(4):626-33.
9. Shaiegan M, Rasouli M, Zadsar M, et al. Meta-analysis of cytomegalovirus seroprevalence in volunteer blood donors and healthy subjects in Iran from 1992 to 2013. Iran J Basic Med Sci 2015; 18(7):627-34
10. Harari A, Zimmerli SC, Pantaleo G. Cytomegalovirus (CMV)-specific cellular immune responses. Hum Immunol 2004; 65: 500-6.
11. Rump K, Rahmel T, Rustige AM, et al. The aquaporin 3 promoter polymorphism− 1431 A/G is associated with acute graft rejection and cytomegalovirus infection in kidney recipients due to altered immune cell migration. Cells 2020; 9: 1421.
12. Shoup M, Ourahmane A, Ginsburg EP, et al. Substitution-inert polynuclear platinum compounds inhibit human cytomegalovirus attachment and entry. Antivir Res 2020; 184:104957
13. Liu Z, Yu Y, Liu H, et al. Establishment of a sandwich light-initiated chemiluminescence assay with double antigen for detecting human cytomegalovirus IgG antibody. J Med Virol 2021; 93:5025-32
14. Awadh N, Nyawale H, Chibwe E, et al. Serological markers of viral infections (rubella virus, human cytomegalovirus and arboviruses) among symptomatic pregnant women in rural and urban areas of Mwanza, Tanzania. Trop Med Infec Dis 2021; 6: 186-212.
15. Zavaglio F, Fiorina L, Suárez NM, et al. Detection of genotype-specific antibody responses to glycoproteins b and h in primary and non-primary human cytomegalovirus infections by peptide-based ELISA. Viruses 2021; 13:399-405.
16. Milavec M, Pavšiˇc J, Košir AB, et al. The performance of human cytomegalovirus digital PCR Reference Measurement Procedure in seven External Quality Assessment schemes over four years. Methods 2021; 25:107-12.
17. Lazar K, Rabe T, Goelz R, et al. Human cytomegalovirus reactivation during lactation: Impact of antibody kinetics and neutralization in blood and breast milk. Nutrients 2020; 12:338-41.
18. Hirsch HH, Lautenschlager I, Pinsky BA, et al. An international multicenter performance analysis of cytomegalovirus load tests. Clin Infect Dis 2013; 56(3):367-73.
19. Ziyaeyan M, Sabahi F. Human cytomegalovirus: infections and diagnosis. Iran J Med Sci 2008; 33(3):127-32.
20. Razonable RR, Humar A. Cytomegalovirus in solid organ transplant recipients- Guidelines of the american society of transplantation infectious diseases community of practice. Clin Transplant 2019; 33(9):e13512.
21. Eshaghi F. Bashi Zadeh Fakhar H, Ghane M, et al. Diagnostic evaluation of Streptococcus gallolyticus infection in patients with colon diseases by polymerase chain reaction (PCR) and culturing methods. Int J Cancer Manag 2020; 13(6):e101903.
22. Merchant EA, Flint K, Barouch DH, et al. Coinfection with coronavirus disease 2019, previously undiagnosed human immunodeficiency virus, Pneumocystis jirovecii pneumonia and cytomegalovirus pneumonitis, with possible immune reconstitution inflammatory syndrome. Idcases 2021; 24:e01153.
23. Covid19.who.int. 2021. WHO Coronavirus (COVID-19). [online].
24. Seyed-Alinaghi S, Afsahi AM, MohsseniPour M, et al. Late complications of COVID-19; a systematic review of current evidence. Arch Acad Emerg Med 2021; 9(1):e14.
25. Kotton NC, Kumar D, Caliendo AM, et al. Updated international consensus guidelines on the management of cytomegalovirus in solidorgan transplantation. Transplantation 2013; 96(4):333-60.
26. Mocarski ES, Shenk T, Pass RF. Cytomegalovirus. In: Knipe DM, Howley PM, ed, Fields Virology, Philadelphia, Lippicott Williams&Wilkins. 2007; PP:2701-2772.
27. Kouri V, Resik S, Enamorado A, et al. Longitudinal study of herpesviruses in kidney transplant recipients in Cuba. Clin Infect Dis 2003; 36(6):818-21.
28. Erice A. Resistance of human cytomegalovirus to antiviral drugs. Clin Microbiol Rev 1999; 12(2): 286-97.
29. Forman SJ, Zaia JA. Treatment and prevention of cytomegalovirus pneumonia after bone marrow transplantation: where do we stand? Blood 1994; 83:2392-8.
30. Enders G, Daiminger A, Lindemann L, et al. Cytomegalovirus (CMV) seroprevalence in pregnant women, bone marrowdonors and adolescents in Germany, 1996-2010. Med Microbiol Immunol 2012; 201(3):303-9
31. Pembrey L, Raynor P, Griffiths P, et al. Seroprevalence of cytomegalovirus, epstein barr virus and varicella zoster virus among pregnant women in bradford: a cohort study. PLoS One 2013; 8(11):e81881.
32. Abu-Madi MA, Behnke JM, Dabritz HA. Toxoplasma gondii seropositivity and co-infection with TORCH pathogens in high-risk patients from Qatar. Am J Trop Med Hyg 2010; 82(4):626-33.
33. Ali S, Khan FA, Mian AA, et al. Seroprevalence of cytomegalovirus, herpes simplex virus and rubella virus among pregnant women in KPK province of Pakistan. J Infect Dev Ctries 2014; 8(3):389-90.
34. Shaiegan M, Rasouli M, Zadsar M, et al. Meta‐analysis of cytomegalovirus sero-prevalence in volunteer blood donors and healthy subjects in Iran from 1992 to 2013. Iran J Basic Med Sci 2015; 18(7):627-34.
35. Motamedifar M, Hashemizadeh Z, Hadi N, et al. The study of seroprevalence of human cytomegalovirus in healthy blood donors referring to Fars blood bank. Hormozgan Med J 2009; 12(4):237-42.[In Persian].
36. Case IPTA. 237 coinfection with COVID-19 and cytomegalovirus in kidney transplant recipients–a case series. Transplant proceed 2021; 54(12):251-260.
37. Bal Z, Uyar M, Tutal E, et al. Cytomegalovirus Infectıon in renal transplant recipients: one center's experience. Transplant proceed 2013; 45(10):3520-3.
38. Chiasakul T, Townamchai N, Jutivorakool K, et al. Risk factors of cytomegalovirus disease in kidney transplant recipients: a single-center study in thailand. Transplant Proceed 2015; 47(8):2460-4.
39. Kalkan IH, Dağli U. What is the most accurate method for the diagnosis of cytomegalovirus (CMV) enteritis or colitis. Turk J Gastroenterol 2010; 21:83-6.
40. Jamalidoust M, Namayandeh M, Pouladfar G, et al. Post-Liver transplant cytomegalovirus (cmv) reactivation, graft, and patient survival rates in iranian population. Jundishapur J Microbiol 2021; 14(3):e112775.
41. Lakzayi E, Fozouni L, Pordeli H, et al. A Seromolecular Study on the prevalence of cytomegalovirus in pregnant women living in Golestan Province, Iran. Shiraz E-Med J 2020; 21(7):e96238.
42. Rahimzadeh G, Shahbaznejad L, Fahimzad A, et al. Seroepidemiology of cytomegalovirus in pregnant women in sari, Iran 2018-2020. J Mazandaran Univ Med Sci 2021; 31(202):149-154.
43. Fadhil HY. Correlation the cytomegalovirus (cmv) igm antibodies and viral dna presence with rheumatoid arthritis (ra) in iraqi patients. Iraqi J Sci 2019; 60(11):2383-2389.
44. Mohsin IH, Badday Hassan I, Daim Saleh MA. A comparative study of immunological and molecular techniques to diagnose human cytomegalovirus in renal failure patients in diyala governor. Ind J Forensic Med Toxicol 2022; 16(1):107-12.
45. Bahri T. Comparison of ELISA, antigenemia assay and nested pcr monitoring techniques for detection of cytomegalovirus infection. J Med Microb Diagn 2018, 7:1.
46. Miller H, Simpson P, Forman M, et al. A novel multiplexed enzyme-linked immunosorbent assay for the detection of igg seroreactivity to cytomegalovirus (CMV) UL144. J Clin Microbiol 2021; 59(8):e00964-21.
47. Shkhair H, Al-janabi O, Abed Kelkawi AH, et al. Comparative evaluation study of ELISA system and MINI-VIDAS system for detection of Cytomegalovirus IgM antibodies. J Pharm Sci Res 2018; 10(10):2549-2550.
48. Çolak D, Sa ̆glık ̇I, Mutlu D, et al. Comparison of two commercial quantitative cy-tomegalovirus (cmv) polymerase chain reaction tests calibrated by world health organization international cmv standard. Mikrobiyoloji Bul 2020; 54:257-265.
49. Stilp AC, König P, Scherer M, et al. Stable and inducible gene knockdown in primary human fibroblasts: a versatile tool to study the role of human cytomegalovirus host cell factors. In Human Cytomegaloviruses; Humana: New York,NY, USA.2021; pp. 115–132.
50. Aghajanian S, Rangani A, Alizadeh-Navaei R, et al. Epidemiology of cytomegalovirus among patients undergoing hemodialysis in Sari, Iran 2017-2018. J Mazandaran Univ Med Sci 2021, 31(199):179-185.
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| Issue | Vol 12 No 4 (2024) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/jmb.v12i4.17006 | |
| Keywords | ||
| Cytomegalovirus Diagnostic ELISA Infection Real time PCR. | ||
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How to Cite
1.
Bali Chalandar F, Bashi Zadeh Fakhar H, Rezaei F. Comparison of RT.PCR and ELISA Methods in the Diagnosis of Human Cytomegalovirus in Kidney Transplant Patients. J Med Bacteriol. 2024;12(4):44-54.
