González-Calixto C, Ruiz-Tachiquín ME, Burgueño-Ferreira J, Aguilera P, Espinoza-Rojo M

Language: Spanish
References: 32
Page: 129-136
PDF size: 56.48 Kb.

ABSTRACT

Early detection of human cytomegalovirus can prevent serious health consequences for immunocompromised patients. Therefore, it is important to employ methods of sensitive and accurate detection. In this paper, we compared the sensitivity and specificity of detection of three methods: 1) COBAS Amplicor ™ CMV MONITOR Test (CACM), commercial and validated method; it uses polymerase chain reaction (PCR) to detect the early gene Pol, 2) real-time PCR, and 3) real-time reverse transcription coupled to PCR (RT-PCR); the 2 and 3 were homemade, which validated the detection of immediate early gene Ie1. We performed a cross-sectional study with 42 positive HIV-1 human plasma samples. Nucleic acids were extracted using two methods: 1) a caotropic agent and 2) an affinity column. We obtained a 9.5, 26.2, and 33.3% of positive samples with CACM, PCR, and RT-PCR, respectively. Using the techniques homemade, we detected a minimum of 26 copies/mL. We used the kappa test to determine correlation between methods, obtaining values of 0.34 (PCR vs CACM), 0.0079 (RT-PCR vs CACM), and 0.65 (RT-PCR vs PCR). The greater sensitivity and specificity were obtained by real time PCR, as well as the largest positive and negative predictive values. We concluded that the qualitative detection of immediate early gene Ie1 (synonymous of active infection) of HCMV in positive HIV-1 human plasma samples by real time PCR could be used as an alternative method to CACM.

REFERENCES

1. Chee MS, Bankier AT, Beck S, Bohni R, Brown CM, Cerny R, et al. Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol. 1990; 154: 125-69.

2. Emery V. Investigation of CMV disease in immunocompromised patients. J Clin Pathol. 2001; 54: 84-8.

3. Mecarski ES, Courcelle CT. Cytomegalovirus and their replication. In: Knipe DMP, Howley M, Griffin DE, Lamb RA, Martin MA, Roizman B, Straus SE, (ed.) Lippincott-Raven; Philadelphia, Pa: Fields Virology; 2001: 2629-73.

4. Luliano R, Forastieri G, Brizzi M, Mecocci L, Mezzotta F, Ceccherin-Nellil. Correlation between plasma HIV-1 RNA levels and the rate of immunologic decline. J Acquir Immune Defic Syndr Hum Retrovirol. 1997; 14: 408-14.

5. Deayton J. Changing trends in cytomegalovirus disease in HIV-infected patients. Herpes. 2001; 8: 2.

6. Centers for disease control and prevention. Revised classification system for HIV infection and expanded surveillance case definition for AIDS among adolescents and adults. Morbid Mortal Weekly Rep. 1993; 41: 1-19.

7. Sainz B, LaMarca H, Garry R, Morris C. Synergistic inhibition of human cytomegalovirus replication by interferon-alfa/beta and interferon-gamma. BMC Virol J. 2005; 2: 14.

8. 8 García-Ramirez J, Ruchti J, Huang H, Simmen K, Angulo A, Ghazal P. Dominance of virus over host factors in cross-species activation of human cytomegalovirus early gene expression. J Virol. 2001; 75: 26-35.

9. Grzimek N, Dreis D, Schmalz S, Reddehase M. Random, asynchronous, and asymmetric transcriptional activity of enhancer-flanking major immediate-early genes ie1/3 and ie2 during murine cytomegalovirus latency in the lungs. J Virol. 2001; 75: 2692-705.

10. Wattamano P, Clayton J, Kopicko J, Kissinger P, Elliot S, Jarrott C, et al. Comparison of three assays for cytomegalovirus detection in AIDS patients at risk for retinitis. J Clin Microbiol. 2000; 38: 727-32.

11. Pang X, Chui L, Featon J, LeBlanc B, Preiksaitis J. Comparison of lightcycler-based PCR, COBAS Amplicor CMV Monitor, and pp65 antigenemia assay for quantitative measurement of cytomegalovirus viral load in peripheral blood specimens from patients after solid organ transplantation. J Clin Microbiol. 2003; 41: 3167-74.

12. Pellegrin I, Garrigue I, Binquet C, Chene G, Neau D, Bonot P, et al. Evaluation of new quantitative assays for diagnosis and monitoring of cytomegalovirus disease in human immunodeficiency virus-positive patients. J Clin Microbiol. 1999; 37: 3124-32.

13. Amorim ML, Cabeda JM, Seca R, Mendes AC, Castro AP, Amorim JM. CMV infection of liver transplant recipients: comparison of antigenemia and molecular biology assays. BMC Infect Dis. 2001; 1: 2.

14. Preiser W, Rabenau H, Vogel J, Brixner V, Doerr H. Performance characteristics of an automated PCR assay for the quantification of cytomegalovirus DNA in plasma. J Virol Microbiol. 2002; 101: 149-57.

15. Najioullah F, Thouvenot D, Lina B. Development of a real-time PCR procedure including an internal control for the measurement of HCMV viral load. J Virol Methods. 2001; 92: 55-64.

16. Onishi Y, Mori S, Higuchi A, Kim S, Fukuda T, Heike Y, et al. Early detection of plasma cytomegalovirus DNA by real-time PCR after allogeneic hematopoietic stem cell transplantation. Tohoku J Exp Med. 2006; 210: 125-35.

17. Yerly S, Perrin L, Van-Delden C, Schaffer S, Thamm S, Wunderli W, Kaiser L. Cytomegalovirus quantification in plasma by an automated real-time PCR assay. J Clin Virol. 2007; 38: 298-303.

18. Gault E, Michel Y, Dehée A, Belabani C, Nicolas J, Garbarg-Chenon A. Quantification of human cytomegalovirus DNA by real-time PCR. J Clin Microbiol. 2001; 37: 772-5.

19. Griscelli F, Barrois M, Chauvin S, Lastere S, Bellet D, Bourhis J. Quantification of human cytomegalovirus DNA in bone marrow transplant recipients by real-time PCR. J Clin Microbiol. 2001; 39: 4362-9.

20. Kalpoe J, Kroes A, De Jong M, Schinkel J, De Brouwer C, Beersma M, et al. Validation of clinical application of cytomegalovirus plasma DNA load measurement and definition of treatment criteria by analysis of correlation to antigen detection. J Clin Microbiol. 2004; 42: 1498-504.

21. Caliendo A, Ingersoll J, Fox-Canele A, Pargman S, Bythwood T, Hayden M, et al. Evaluation of real-time PCR laboratory-developed test using analyte-specific reagents for cytomegalovirus quantification. J Clin Microbiol. 2007; 45: 1723-7.

22. Nolan T, Hands R, Bustin S. Quantification of mRNA using real-time RT-PCR. Nat Protoc. 2006; 1: 1559-82.

23. Machida U. Kami M, Fukui T, Kazuyama Y, Kinoshita M, Tanaka Y, et al. Real-Time automated PCR for early diagnosis and monitoring of cytomegalovirus infection after bone marrow transplantation. J Clin Microbiol. 2000; 3: 2536-42.

24. Gerna G, Baldanti F, Lilleri D, Parea M, Alessandrino E, Pagani A, Locatelli F, et al. Human cytomegalovirus immediate-early mRNA detection by nucleic acid sequence-based amplification as a new parameter for preemptive therapy in bone marrow transplant recipients. J Clin Microbiol. 2000; 38: 1845-53.

25. Roche Molecular Systems, Inc. COBAS AMPLICOR CMV MONITORTM Test, Roche Diagnostics. P/N: 03563898 190. Rev. 2003; 1:1-38.

26. Cohen J. A coefficient of agreement for nominal scales. Educ Psychol Measur. 1960; 20: 37-46.

27. Fleiss JL, Cohen J, Everitt BS. Large-sample standard errors of kappa and weighted kappa. Psychological Bulletin. 1969: 72: 323-7.

28. Fleiss JL. Statistical methods for rates and proportions. Second Edition, New York: John Wiley & Sons, Inc; 1981.

29. Duffau TG, Navarrete RL, Fernández CC. Estudio de concordancia clínica en educandos de pre y postítulo en pediatría. Rev Chil Pediatr. 2000; 71: 340-6.

30. Schaade L, Kockelkorn P, Ritter K, Kleines M. Detection of cytomegalovirus DNA in human specimens by light cycler PCR. J Clin Microbiol. 2000; 38: 4006-9.

31. Herrmann B, Larsson V, Rubin C, Sund F, Eriksson B, Arvidson J, et al. Comparison of a duplex quantitative real-time PCR assay and the COBAS Amplicor CMV Monitor test for detection of cytomegalovirus. J Clin Microbiol. 2004; 42: 1909-14.

32. Boom R, Sol C, Schuurman T, Breda A, Weel J, Beld M, et al. Human cytomegalovirus DNA in plasma and serum specimens of renal transplant recipients is highly fragmented. J Clin Microbiol. 2002; 40: 4105-13.