Evaluation of ELISA and Real-Time PCR Results Sent for The Diagnosis of Cytomegalovirus (CMV) İnfection
Öz
AİM: Human Cytomegalovirus (CMV) is an enveloped virus with a linear double-stranded DNA genome and an icosahedral capsid, belonging to the Betaherpesvirinae subfamily of the Herpesviridae family. While it is rarely pathogenic in immunocompetent individuals, CMV poses a significant health threat to immunocompromised patients. Due to the strong association between CMV infection and decreased patient survival, the monitoring and treatment of CMV infection are crucial. In immunosuppressed individuals, serological test results have significant limitations in diagnosing acute CMV infection; in such cases, the detection of viral nucleic acid, most commonly by polymerase chain reaction (PCR), is used to confirm the presence of CMV. There are limited studies investigating the correlation between these two testing methods. Therefore, this study aims to comparatively evaluate CMV antibody and CMV DNA test results.
MATERIAL AND METHODS: The Bosphore CMV quantification kit was used in the PCR phase. CMV-IgM and CMV-IgG antibody levels were analyzed using the ELISA method (Cobas E 411/Roche) according to the procedure described by the manufacturer.
RESULTS: In 81% of the samples (298/367), the results are consistent; both test results were negative in 280 (76.2%) samples and positive in 18 (4.9%) samples. It was found that 92.7% of the 302 patients who tested negative for IgM were also negative for CMV-PCR. Only 45% of the 40 patients who tested positive for CMV-PCR were identified as positive by the IgM test. Statistical analysis shows that there is a significant difference in the detection rates of this pathogen between the two tests (p < 0.001).
CONCLUSIONS: There is a strong concordance between the negative results of both tests; however, the same concordance has not been observed among positive results. Overall, further studies involving larger patient cohorts are recommended to improve CMV management strategies in clinical practice.
Kaynakça
- KAYNAKLAR 1. Mocarski ES, Courcelle CT. Cytomegaloviruses and their replication. Fields Virology. 4th ed. In: Knipe DM, Howley PM, editors. Philadelphia: Lippincott Williams & Wilkins; 2001:2629–2673.
- 2. Emery VC. Investigation of CMV disease in immunocompromised patients. J Clin Pathol. 2001;54(2):84–88. doi:10.1136/jcp.54.2.84
- 3. Sweet C. The pathogenicity of cytomegalovirus. FEMS Microbiol Rev. 1999;23(4):457–82. doi:10.1111/j.1574-6976.1999.tb00408.x
- 4. Stowell JD, Forlin-Passoni D, Din E, et al. Cytomegalovirus survival on common environmental surfaces: opportunities for viral transmission. J Infect Dis. 2012;205(2):211–4. doi:10.1093/infdis/jir722
- 5. Benz C, Mternohen O, Wulf A, et al. Activated virus-specific T cells are early indicators of anti-CMV immune reaction in liver transplant patients. Gastroenterology. 2000;5:46–122. doi:10.1053/gast.2002.33021
- 6. Minz RW, Kumar M, Kanwar DB, et al. Cytomegalovirus infection in postrenal transplant recipients: 18 years’ experience from a tertiary referral center. Transplant Proc. 2020;52(10):3173–8. doi:10.1016/j.transproceed.2020.08.011
- 7. Razonable RR, Paya CV, Smith TF. Role of the laboratory in diagnosis and management of cytomegalovirus infection in hematopoietic stem cell and solid-organ transplant recipients. J Clin Microbiol. 2002;40(3):746–52. doi:10.1128/JCM.40.3.746-752.2002
- 8. Klemola E, Kääriäinen L. Cytomegalovirus as a possible cause of a disease resembling infectious mononucleosis. Br Med J. 1965;2(5463):1099–102.doi:10.1136/bmj.2.5470.1099
- 9. Bate SL, Dollard SC, Cannon MJ. Cytomegalovirus seroprevalence in the United States: the national health and nutrition examination surveys, 1988–2004. Clin Infect Dis. 2010;50(11):1439–47. doi:10.1086/652438
- 10. Lanzieri TM, Kruszon-Moran D, Amin MM, et al. Seroprevalence of cytomegalovirus among children 1 to 5 years of age in the United States from the National Health and Nutrition Examination Survey of 2011 to 2012. Clin Vaccine Immunol. 2015;22(3):245–7. doi:10.1128/CVI.00697-14
- 11. Lazzarotto T, Spezzacatena P, Pradelli P, Abate DA, Varani S, Landini MP. Avidity of immunoglobulin G directed against human cytomegalovirus during primary and secondary infections in immunocompetent and immunocompromised subjects. Clin Diagn Lab Immunol. 1997;4(4):469–73. doi:10.1128/cdli.4.4.469-473.1997
- 12. Bozdemir T, Daloğlu AE, Çolak D, Gürkan N, Öngüt G, Mutlu D. Tam otomatize sistemlerde çalışılan sitomegalovirüs polimeraz zincir reaksiyonu test sonuçlarının karşılaştırılması: Roche Cobas 6800 ile Qiagen NeuMoDx deneyimi. Mikrobiyol Bul. 2024;58(3):284–92. doi:10.5578/mb.20249727
- 13. Limaye AP, Babu TM, Boeckh M. Progress and challenges in the prevention, diagnosis, and management of cytomegalovirus infection in transplantation. Clin Microbiol Rev. 2021;34(1):e00043-19. doi:10.1128/CMR.00043-19
- 14. Boppana SB, Ross SA, Shimamura M, et al. Saliva polymerase-chain-reaction assay for cytomegalovirus screening in newborns. N Engl J Med. 2011;364(22):2111-2118. doi:10.1056/NEJMoa1006561
- 15. Enan KA, Rennert H, El-Eragi AM, El Hussein ARM, Elkhidir IM. Comparison of Real-time PCR to ELISA for the detection of human cytomegalovirus infection in renal transplant patients in the Sudan. Virol J. 2011;8:222. doi:10.1186/1743-422X-8-222
- 16. Heli P. Quantitative PCR in the diagnosis and monitoring of human cytomegalovirus infection in organ transplant patients. PhD Thesis. Helsinki: University of Helsinki; 2004. p. 55–70.
- 17. Kotton CN. CMV: prevention, diagnosis and therapy. Am J Transplant. 2013;13(Suppl 3):24-40. doi:10.1111/ajt.12006
- 18. Özkarataş E, Özbek ÖA, Oğuz V, Sayiner AA. Solid organ nakli alıcılarında CMV antijenemi testi ve CMV-DNA PCR sonuçlarının karşılaştırılması. Mikrobiyol Bul. 2016;50(1):44–52. doi:10.5578/mb.10701
- 19. Dioverti MV, Razonable RR. Cytomegalovirus. Microbiol Spectrum. 2016;4(4):DMIH2-0022-2015. doi:10.1128/microbiolspec.DMIH2-0022-2015
- 20. Larsson S, Söderberg-Naucler C, Wang FZ, Moller E. Cytomegalovirus DNA can be detected in peripheral blood mononuclear cells from all seropositive and most seronegative healthy blood donors over time. Transfusion. 1998;38(3):271–8. doi:10.1046/j.1537-2995.1998.38398222864.x
- 21. Paya CV, Wilson JA, Espy MJ, et al. Preemptive use of oral ganciclovir to prevent cytomegalovirus infection in liver transplant patients: a randomized, placebo-controlled trial. J Infect Dis. 2002;185(6):854–60. doi:10.1086/339342
- 22. Naumnik B, Małyszko J, Chyczewski L, Kovalchuk O, Małyszko J, Myśliwiec M. Comparison of serology assays and polymerase chain reaction for the monitoring of active cytomegalovirus infection in renal transplant recipients. Transplant Proc. 2007;39(9):2748–50. doi:10.1016/j.transproceed.2007.08.040
- 23. Revello MG, Gerna G. Diagnosis and management of human cytomegalovirus infection in the mother, fetus, and newborn infant. Clin Microbiol Rev. 2002;15(4):680-715. doi:10.1128/CMR.15.4.680-715.2002
- 24. Hodson EM, Jones CA, Webster AC, et al. Antiviral medications to prevent cytomegalovirus disease and early death in recipients of solid-organ transplants: a systematic review of randomised controlled trials. Lancet. 2005;365(9477):2105-2116. doi:10.1016/S0140-6736(05)66553-1
- 25. Bozdemir T, Daloğlu AE, Çolak D, ve ark. Comparison of CMV PCR results between Roche Cobas 6800 and Qiagen NeuMoDx systems. Mikrobiyol Bul. 2024;58(3):284-292. doi:10.5578/mb.20249727
Cytomegalovırus (CMV) Enfeksiyonu Tanısı İçin Gönderilen ELISA ve Real Time PCR Sonuçlarının Değerlendirilmesi
Öz
AMAÇ: İnsan Cytomegalovirus (CMV), Herpesviridae ailesinin Betaherpesvirinae alt familyasına ait lineer çift iplikli, ikozahedral kapsidli zarflı bir virüstür. İmmünokompetan bireylerde nadiren patojenik olmasına rağmen, virüs immünokompromize bireyler için önemli bir sağlık tehdidi oluşturmaktadır. CMV enfeksiyonu ile hasta sağ kalımındaki azalma arasındaki anlamlı ilişki nedeniyle CMV enfeksiyonunun takip ve tedavisi çok önemlidir. Bağışıklık sistemi baskılanmış bireylerde akut CMV enfeksiyonunun teşhisi için serolojik test sonuçlarıyla hastaların değerlendirilmesinde önemli sınırlamalar bulunmaktadır; bu durumda viral nükleik asit tespiti en yaygın olarak polimeraz zincir reaksiyonu (PCR) ile CMV’ nin varlığı ortaya konabilir. Bu iki testin korelasyon düzeyi ile ilgili yapılan çok fazla sayıda çalışma yoktur; bu sebeple bu çalışmada CMV antikorlarının ve CMV DNA sonuçlarının karşılaştırmalı olarak değerlendirilmesini hedeflenmiştir.
GEREÇ ve YÖNTEM: PCR aşamasında Bosphore CMV kantifikasyon kiti kullanılmıştır. CMV-IgM ve CMV-IgG antikor düzeyleri makro ELISA yöntemiyle (Cobas E 411/Roche) üretici tarafından açıklanan prosedüre göre çalışılmıştır.
BULGULAR: Örneklerin %81 (298/367)’inde sonuçlar uyumludur; her iki test sonucu 280 (%76,2) örnekte negatif, 18 (%4,9) örnekte pozitif saptandı. IgM için negatif çıkan 302 hastanın %92,7'sinin CMV-PCR için de negatif olduğu ortaya konmuştur. 40 CMV-PCR pozitif hastanın yalnızca %45'i IgM testiyle pozitif olarak tanımlandı. İstatistiksel analiz, iki test arasında bu patojeni saptama oranları arasında anlamlı bir fark olduğunu göstermektedir (p<0,001).
SONUÇ: Her iki testin negatif sonuçları arasında güçlü bir uyum vardır. Aynı uyum pozitif sonuçlar arasında gözlemlenememiştir. Genel olarak, klinik uygulamalarda CMV yönetim stratejilerini geliştirmek amacıyla daha geniş hasta gruplarıyla yapılacak ileri araştırmalar önerilmektedir.
Kaynakça
- KAYNAKLAR 1. Mocarski ES, Courcelle CT. Cytomegaloviruses and their replication. Fields Virology. 4th ed. In: Knipe DM, Howley PM, editors. Philadelphia: Lippincott Williams & Wilkins; 2001:2629–2673.
- 2. Emery VC. Investigation of CMV disease in immunocompromised patients. J Clin Pathol. 2001;54(2):84–88. doi:10.1136/jcp.54.2.84
- 3. Sweet C. The pathogenicity of cytomegalovirus. FEMS Microbiol Rev. 1999;23(4):457–82. doi:10.1111/j.1574-6976.1999.tb00408.x
- 4. Stowell JD, Forlin-Passoni D, Din E, et al. Cytomegalovirus survival on common environmental surfaces: opportunities for viral transmission. J Infect Dis. 2012;205(2):211–4. doi:10.1093/infdis/jir722
- 5. Benz C, Mternohen O, Wulf A, et al. Activated virus-specific T cells are early indicators of anti-CMV immune reaction in liver transplant patients. Gastroenterology. 2000;5:46–122. doi:10.1053/gast.2002.33021
- 6. Minz RW, Kumar M, Kanwar DB, et al. Cytomegalovirus infection in postrenal transplant recipients: 18 years’ experience from a tertiary referral center. Transplant Proc. 2020;52(10):3173–8. doi:10.1016/j.transproceed.2020.08.011
- 7. Razonable RR, Paya CV, Smith TF. Role of the laboratory in diagnosis and management of cytomegalovirus infection in hematopoietic stem cell and solid-organ transplant recipients. J Clin Microbiol. 2002;40(3):746–52. doi:10.1128/JCM.40.3.746-752.2002
- 8. Klemola E, Kääriäinen L. Cytomegalovirus as a possible cause of a disease resembling infectious mononucleosis. Br Med J. 1965;2(5463):1099–102.doi:10.1136/bmj.2.5470.1099
- 9. Bate SL, Dollard SC, Cannon MJ. Cytomegalovirus seroprevalence in the United States: the national health and nutrition examination surveys, 1988–2004. Clin Infect Dis. 2010;50(11):1439–47. doi:10.1086/652438
- 10. Lanzieri TM, Kruszon-Moran D, Amin MM, et al. Seroprevalence of cytomegalovirus among children 1 to 5 years of age in the United States from the National Health and Nutrition Examination Survey of 2011 to 2012. Clin Vaccine Immunol. 2015;22(3):245–7. doi:10.1128/CVI.00697-14
- 11. Lazzarotto T, Spezzacatena P, Pradelli P, Abate DA, Varani S, Landini MP. Avidity of immunoglobulin G directed against human cytomegalovirus during primary and secondary infections in immunocompetent and immunocompromised subjects. Clin Diagn Lab Immunol. 1997;4(4):469–73. doi:10.1128/cdli.4.4.469-473.1997
- 12. Bozdemir T, Daloğlu AE, Çolak D, Gürkan N, Öngüt G, Mutlu D. Tam otomatize sistemlerde çalışılan sitomegalovirüs polimeraz zincir reaksiyonu test sonuçlarının karşılaştırılması: Roche Cobas 6800 ile Qiagen NeuMoDx deneyimi. Mikrobiyol Bul. 2024;58(3):284–92. doi:10.5578/mb.20249727
- 13. Limaye AP, Babu TM, Boeckh M. Progress and challenges in the prevention, diagnosis, and management of cytomegalovirus infection in transplantation. Clin Microbiol Rev. 2021;34(1):e00043-19. doi:10.1128/CMR.00043-19
- 14. Boppana SB, Ross SA, Shimamura M, et al. Saliva polymerase-chain-reaction assay for cytomegalovirus screening in newborns. N Engl J Med. 2011;364(22):2111-2118. doi:10.1056/NEJMoa1006561
- 15. Enan KA, Rennert H, El-Eragi AM, El Hussein ARM, Elkhidir IM. Comparison of Real-time PCR to ELISA for the detection of human cytomegalovirus infection in renal transplant patients in the Sudan. Virol J. 2011;8:222. doi:10.1186/1743-422X-8-222
- 16. Heli P. Quantitative PCR in the diagnosis and monitoring of human cytomegalovirus infection in organ transplant patients. PhD Thesis. Helsinki: University of Helsinki; 2004. p. 55–70.
- 17. Kotton CN. CMV: prevention, diagnosis and therapy. Am J Transplant. 2013;13(Suppl 3):24-40. doi:10.1111/ajt.12006
- 18. Özkarataş E, Özbek ÖA, Oğuz V, Sayiner AA. Solid organ nakli alıcılarında CMV antijenemi testi ve CMV-DNA PCR sonuçlarının karşılaştırılması. Mikrobiyol Bul. 2016;50(1):44–52. doi:10.5578/mb.10701
- 19. Dioverti MV, Razonable RR. Cytomegalovirus. Microbiol Spectrum. 2016;4(4):DMIH2-0022-2015. doi:10.1128/microbiolspec.DMIH2-0022-2015
- 20. Larsson S, Söderberg-Naucler C, Wang FZ, Moller E. Cytomegalovirus DNA can be detected in peripheral blood mononuclear cells from all seropositive and most seronegative healthy blood donors over time. Transfusion. 1998;38(3):271–8. doi:10.1046/j.1537-2995.1998.38398222864.x
- 21. Paya CV, Wilson JA, Espy MJ, et al. Preemptive use of oral ganciclovir to prevent cytomegalovirus infection in liver transplant patients: a randomized, placebo-controlled trial. J Infect Dis. 2002;185(6):854–60. doi:10.1086/339342
- 22. Naumnik B, Małyszko J, Chyczewski L, Kovalchuk O, Małyszko J, Myśliwiec M. Comparison of serology assays and polymerase chain reaction for the monitoring of active cytomegalovirus infection in renal transplant recipients. Transplant Proc. 2007;39(9):2748–50. doi:10.1016/j.transproceed.2007.08.040
- 23. Revello MG, Gerna G. Diagnosis and management of human cytomegalovirus infection in the mother, fetus, and newborn infant. Clin Microbiol Rev. 2002;15(4):680-715. doi:10.1128/CMR.15.4.680-715.2002
- 24. Hodson EM, Jones CA, Webster AC, et al. Antiviral medications to prevent cytomegalovirus disease and early death in recipients of solid-organ transplants: a systematic review of randomised controlled trials. Lancet. 2005;365(9477):2105-2116. doi:10.1016/S0140-6736(05)66553-1
- 25. Bozdemir T, Daloğlu AE, Çolak D, ve ark. Comparison of CMV PCR results between Roche Cobas 6800 and Qiagen NeuMoDx systems. Mikrobiyol Bul. 2024;58(3):284-292. doi:10.5578/mb.20249727
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Viroloji |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Erken Görünüm Tarihi | 9 Mayıs 2025 |
| Yayımlanma Tarihi | 2 Mayıs 2025 |
| Gönderilme Tarihi | 6 Şubat 2025 |
| Kabul Tarihi | 21 Mart 2025 |
| Yayımlandığı Sayı | Yıl 2025 Cilt: 9 Sayı: 1 |
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