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Comparison of quantitative cytomegalovirus real-time PCR in whole blood and pp65 antigenemia assay: clinical utility of CMV real-time PCR in hematopoietic stem cell transplant recipients.

Choi SM, Lee DG, Lim J, Park SH, Choi JH, Yoo JH, Lee JW, Kim Y, Han K, Min WS, Shin WS, Kim CC - J. Korean Med. Sci. (2009)

Bottom Line: Exposure to the antiviral agent influenced the results of the two assays.Via ROC curve analysis, the tentative cutoff value for preemptive therapy was determined to be approximately 2x10(4) copies/mL (sensitivity, 80.0%; specificity, 50.0%) in the high risk patients, and approximately 3x10(4) copies/mL (sensitivity, 90.0%; specificity, 70.0%) in the patients at low risk for CMV disease.Further study to validate the optimal cutoff value for the initiation of preemptive therapy is currently underway.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, The Catholic Hemopoietic Stem Cell Transplantation Center, College of Medicine, The Catholic University of Korea, Seoul, Korea.

ABSTRACT
Successful preemptive therapy for cytomegalovirus (CMV) infection in transplant patients depends on the availability of sensitive, specific, and timely diagnostic tests for CMV infection. Although the pp65 antigenemia assay has been widely used for this purpose, real-time quantification of CMV DNA has recently been recognized as an alternative diagnostic approach. However, the guidelines for antiviral therapy based on real-time quantitative polymerase chain reaction (RQ-PCR) have yet to be established. From November 2004 to March 2005, a total of 555 whole blood samples from 131 hematopoietic stem cell transplant (HSCT) recipients were prospectively collected. RQ-PCR was conducted using an Artus CMV LC PCR kit (QIAGEN). Both qualitative and quantitative correlations were drawn between the two methods. Exposure to the antiviral agent influenced the results of the two assays. Additionally, the discrepancy was observed at low levels of antigenemia and CMV DNA load. Via ROC curve analysis, the tentative cutoff value for preemptive therapy was determined to be approximately 2x10(4) copies/mL (sensitivity, 80.0%; specificity, 50.0%) in the high risk patients, and approximately 3x10(4) copies/mL (sensitivity, 90.0%; specificity, 70.0%) in the patients at low risk for CMV disease. Further study to validate the optimal cutoff value for the initiation of preemptive therapy is currently underway.

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Longitudinal trend of antigenemia values and CMV DNA load in two patients (A, B) who developed CMV diseases. The details are provided in the text.CMV, cytomegalovirus; CTL, cytotoxic T lymphocyte; FOS, foscarnet; GCV, ganciclovir; RQ-PCR, real-time quantitative polymerase chain reaction.
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Figure 3: Longitudinal trend of antigenemia values and CMV DNA load in two patients (A, B) who developed CMV diseases. The details are provided in the text.CMV, cytomegalovirus; CTL, cytotoxic T lymphocyte; FOS, foscarnet; GCV, ganciclovir; RQ-PCR, real-time quantitative polymerase chain reaction.

Mentions: Upon longitudinal analysis for the individual patients, the follow-up curves of the CMV DNA load by RQ-PCR and pp65 antigenemia were uniform and no discrepancies were noted between the two methods in response to anti-CMV therapy (Fig. 2). The duration of follow up of enrolled patients was a median of 1,126 days with a range of 22-3,099 days. There were 2 confirmed cases of CMV diseases (Fig. 3). Patient A was a 7-yr old girl who had undergone cord blood transplantation and experienced acute GVHD (grade IV). CMV infection was detected by both methods at the 20th day after transplantation and preemptive ganciclovir (GCV) therapy was instituted and both viral copies and antigenemia were decreased to zero. After the cessation of GCV, the viral DNA loads increased earlier than antigenemia and both increased continuously despite the GCV therapy. GCV was switched to foscarnet (FOS) but the patient developed CMV pneumonia and died as a result of it. Patient B was a 57-yr old woman who underwent peripheral blood stem cell transplantation from a matched sibling donor. She received alemtuzumab as a conditioning regimen. CMV infection was detected by both RQ-PCR and pp65 antigenemia on the 21st day after transplantation. Viral load and the number of pp65 antigen-positive cells were somewhat discrepant during the early period of antiviral therapy. She received GCV and FOS with CMV immunoglobulin alone or in combination, guided by pp65 antigenemia. However, CMV retinitis and pneumonia developed. After 2 months of intravenous antiviral therapy, she was treated with maintenance oral GCV and CMV specific cytotoxic T lymphocyte (CTL) immunotherapy once per month. She survived, and the viral load was reduced below detectable levels by both methods.


Comparison of quantitative cytomegalovirus real-time PCR in whole blood and pp65 antigenemia assay: clinical utility of CMV real-time PCR in hematopoietic stem cell transplant recipients.

Choi SM, Lee DG, Lim J, Park SH, Choi JH, Yoo JH, Lee JW, Kim Y, Han K, Min WS, Shin WS, Kim CC - J. Korean Med. Sci. (2009)

Longitudinal trend of antigenemia values and CMV DNA load in two patients (A, B) who developed CMV diseases. The details are provided in the text.CMV, cytomegalovirus; CTL, cytotoxic T lymphocyte; FOS, foscarnet; GCV, ganciclovir; RQ-PCR, real-time quantitative polymerase chain reaction.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2719194&req=5

Figure 3: Longitudinal trend of antigenemia values and CMV DNA load in two patients (A, B) who developed CMV diseases. The details are provided in the text.CMV, cytomegalovirus; CTL, cytotoxic T lymphocyte; FOS, foscarnet; GCV, ganciclovir; RQ-PCR, real-time quantitative polymerase chain reaction.
Mentions: Upon longitudinal analysis for the individual patients, the follow-up curves of the CMV DNA load by RQ-PCR and pp65 antigenemia were uniform and no discrepancies were noted between the two methods in response to anti-CMV therapy (Fig. 2). The duration of follow up of enrolled patients was a median of 1,126 days with a range of 22-3,099 days. There were 2 confirmed cases of CMV diseases (Fig. 3). Patient A was a 7-yr old girl who had undergone cord blood transplantation and experienced acute GVHD (grade IV). CMV infection was detected by both methods at the 20th day after transplantation and preemptive ganciclovir (GCV) therapy was instituted and both viral copies and antigenemia were decreased to zero. After the cessation of GCV, the viral DNA loads increased earlier than antigenemia and both increased continuously despite the GCV therapy. GCV was switched to foscarnet (FOS) but the patient developed CMV pneumonia and died as a result of it. Patient B was a 57-yr old woman who underwent peripheral blood stem cell transplantation from a matched sibling donor. She received alemtuzumab as a conditioning regimen. CMV infection was detected by both RQ-PCR and pp65 antigenemia on the 21st day after transplantation. Viral load and the number of pp65 antigen-positive cells were somewhat discrepant during the early period of antiviral therapy. She received GCV and FOS with CMV immunoglobulin alone or in combination, guided by pp65 antigenemia. However, CMV retinitis and pneumonia developed. After 2 months of intravenous antiviral therapy, she was treated with maintenance oral GCV and CMV specific cytotoxic T lymphocyte (CTL) immunotherapy once per month. She survived, and the viral load was reduced below detectable levels by both methods.

Bottom Line: Exposure to the antiviral agent influenced the results of the two assays.Via ROC curve analysis, the tentative cutoff value for preemptive therapy was determined to be approximately 2x10(4) copies/mL (sensitivity, 80.0%; specificity, 50.0%) in the high risk patients, and approximately 3x10(4) copies/mL (sensitivity, 90.0%; specificity, 70.0%) in the patients at low risk for CMV disease.Further study to validate the optimal cutoff value for the initiation of preemptive therapy is currently underway.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, The Catholic Hemopoietic Stem Cell Transplantation Center, College of Medicine, The Catholic University of Korea, Seoul, Korea.

ABSTRACT
Successful preemptive therapy for cytomegalovirus (CMV) infection in transplant patients depends on the availability of sensitive, specific, and timely diagnostic tests for CMV infection. Although the pp65 antigenemia assay has been widely used for this purpose, real-time quantification of CMV DNA has recently been recognized as an alternative diagnostic approach. However, the guidelines for antiviral therapy based on real-time quantitative polymerase chain reaction (RQ-PCR) have yet to be established. From November 2004 to March 2005, a total of 555 whole blood samples from 131 hematopoietic stem cell transplant (HSCT) recipients were prospectively collected. RQ-PCR was conducted using an Artus CMV LC PCR kit (QIAGEN). Both qualitative and quantitative correlations were drawn between the two methods. Exposure to the antiviral agent influenced the results of the two assays. Additionally, the discrepancy was observed at low levels of antigenemia and CMV DNA load. Via ROC curve analysis, the tentative cutoff value for preemptive therapy was determined to be approximately 2x10(4) copies/mL (sensitivity, 80.0%; specificity, 50.0%) in the high risk patients, and approximately 3x10(4) copies/mL (sensitivity, 90.0%; specificity, 70.0%) in the patients at low risk for CMV disease. Further study to validate the optimal cutoff value for the initiation of preemptive therapy is currently underway.

Show MeSH
Related in: MedlinePlus