**7. Acknowledgement**

The authors thank other members of the laboratory. P.P-R. was funded by Instituto de Salud Carlos III, Programa Miguel Servet CP05/00226.

#### **8. References**

416 Advances in Hematopoietic Stem Cell Research

detection (Caliendo et al., 2009; Ikewaki et al., 2005). Although, the variability in the results obtained with commercial assays (kits or ASRs) are proven to be lower than when using in-

The most critical factor for standardization is the lack of using an universally accepted standard for CMV quantification will make possible the comparison among results thus establishing a common management of patients in different centers (Atkinson & Emery, 2011). The reference material used as a calibrator have to be traceable and commutable to achieve accurate clinical results ensuring consistency with clinical samples (Caliendo et al., 2009). While many laboratories produce their own calibrator as an attempt of standardizing some CMV reference material is under development and in fact there are standards commercially available for quantification. For instance, the National Institute of Standards and Technology (NIST) started the development of a reference standard for CMV based on pure CMV DNA from a Towne strain from which after some modification, the final construct will be used to produce viral DNA (Wang et al., 2004). Moreover, the OptiQuant® CMV DNA Quantification Panel from AcroMetrix has been carefully formulated to mimic naturally occurring human specimens containing CMV viral DNA. It consists of cultured CMV that has been diluted in defibrinated, delipidized normal human plasma at different concentrations. The panel can be used with any test procedure designed for measuring CMV DNA in human serum or plasma and it has been widely used as a standard in several studies (Bravo et al.; Forman et al., 2011; Hanson et al., 2007; Raggam et al., 2010) to compare

In the absence of standardization the current clinical guidelines recommend to each individual laboratory to establish their own viral thresholds for CMV management, (Kotton et al., 2010; Razonable & Emery, 2004), threshold that cover a wide range of viral loads varying from 200-500 copies/mL in some laboratories (Ikewaki et al., 2005; Mori et al., 2002)

In conclusion, the quantitative PCR assays have demonstrated to be more suitable and clinically relevant for the monitoring of the CMV viral load, and the management of patients that develop CMV infection. Although there are several commercially available RT-PCR assays developed for the detection of CMV infection in clinical samples, there are variations in the performance of these tests which limit to establish a common cutoff between different assays. Each laboratory must establish its own cutoff value and monitor clinical outcomes to verify the trigger points used. Using universally accepted standards for CMV quantification will make possible the comparison among results establishing a common management of patients in different centers. However, in the absence of standardization the current clinical guidelines recommend to each individual laboratory to establish their own viral thresholds for CMV management. Further studies are still necessary to establish standardized cut-off

The authors thank other members of the laboratory. P.P-R. was funded by Instituto de Salud

house developed methods (Pang et al., 2009).

techniques with the same laboratory and inter-laboratory.

to 2000-5000 copies/mL in others (Humar et al., 1999).

values in large series of transplant recipients.

Carlos III, Programa Miguel Servet CP05/00226.

**7. Acknowledgement** 

**6. Conclusions** 


Detection of CMV Infection in Allogeneic SCT Recipients: The Multiple Assays 419

Forman, M.; Wilson, A.; & Valsamakis, A. (2011). Cytomegalovirus DNA quantification

Gartner, B. C.; Fischinger, J. M.; Litwicki, A.; Roemer, K.; & Mueller-Lantzsch, N. (2004).

Gault, E.; Michel, Y.; Dehee, A.; Belabani, C.; Nicolas, J. C.; & Garbarg-Chenon, A. (2001).

Gerna, G.; Lilleri, D.; Caldera, D.; Furione, M.; Zenone Bragotti, L.; & Aless&rino, E. P.

Ghisetti, V.; Barbui, A.; Franchello, A.; Varetto, S.; Pittaluga, F.; Bobbio, M.; Salizzoni, M.; &

Gimeno, C.; Solano, C.; Latorre, J. C.; Hern&ez-Boluda, J. C.; Clari, M. A.; Remigia, M. J.;

Gleaves, C. A.; Smith, T. F.; Shuster, E. A.; & Pearson, G. R. (1984). Rapid detection of

Gouarin, S.; Vabret, A.; Scieux, C.; Agbalika, F.; Cherot, J.; Mengelle, C.; Deback, C.;

Gracia-Ahufinger, I.; Tormo, N.; Espigado, I.; Solano, C.; Urbano-Ispizua, A.; Clari, M. A.; de

Griffiths, P. D.; Panjwani, D. D.; Stirk, P. R.; Ball, M. G.; Ganczakowski, M.; Blacklock, H. A.;

Griscelli, F.; Barrois, M.; Chauvin, S.; Lastere, S.; Bellet, D.; & Bourhis, J. H. (2001).

Hakki, M.; Riddell, S. R.; Storek, J.; Carter, R. A.; Stevens-Ayers, T.; Sudour, P.; White, K.;

setup. *J Clin Microbiol* Vol.49, No.7, pp. 2703-5, ISSN 1098-660X

No.8, pp. 3881-2, ISSN 0095-1137

Vol.39, No.2, pp. 772-5, ISSN 0095-1137

Vol.73, No.2, pp. 223-9, ISSN 0146-6615

Vol.46, No.10, pp. 3311-8, ISSN 1098-660X

Vol.19, No.6, pp. 917-9, ISSN 0095-1137

Vol.146, No.1-2, pp. 147-54, ISSN 0166-0934

Vol.48, No.2, pp. 142-6, ISSN 1873-5967

Vol.2, No.8414, pp. 1242-5, ISSN 0140-6736

1137

*Transplant* Vol.41, No.10, pp. 873-9, ISSN 0268-3369

using an automated platform for nucleic Acid extraction & real-time PCR assay

Evaluation of a new automated, st&ardized generic nucleic acid extraction method (total nucleic acid isolation kit) used in combination with cytomegalovirus DNA quantification by COBAS AMPLICOR CMV MONITOR. *J Clin Microbiol* Vol.42,

Quantification of human cytomegalovirus DNA by real-time PCR. *J Clin Microbiol*

(2008). Validation of a DNAemia cutoff for preemptive therapy of cytomegalovirus infection in adult hematopoietic stem cell transplant recipients. *Bone Marrow* 

Marchiaro, G. (2004). Quantitation of cytomegalovirus DNA by the polymerase chain reaction as a predictor of disease in solid organ transplantation. *J Med Virol*

Furio, S.; Calabuig, M.; Tormo, N.; & Navarro, D. (2008). Quantification of DNA in plasma by an automated real-time PCR assay (cytomegalovirus PCR kit) for surveillance of active cytomegalovirus infection and guidance of preemptive therapy for allogeneic hematopoietic stem cell transplant recipients. *J Clin Microbiol*

cytomegalovirus in MRC-5 cells inoculated with urine specimens by using lowspeed centrifugation and monoclonal antibody to an early antigen. *J Clin Microbiol*

Petitjean, J.; Dina, J.; & Freymuth, F. (2007). Multicentric evaluation of a new commercial cytomegalovirus real-time PCR quantitation assay. *J Virol Methods*

la Cruz-Vicente, F.; Navarro, D.; & Perez-Romero, P. (2010). Differences in cytomegalovirus plasma viral loads measured in allogeneic hematopoietic stem cell transplant recipients using two commercial real-time PCR assays. *J Clin Virol*

& Prentice, H. G. (1984). Rapid diagnosis of cytomegalovirus infection in immunocompromised patients by detection of early antigen fluorescent foci. *Lancet*

Quantification of human cytomegalovirus DNA in bone marrow transplant recipients by real-time PCR. *J Clin Microbiol* Vol.39, No.12, pp. 4362-9, ISSN 0095-

Corey, L.; & Boeckh, M. (2003). Immune reconstitution to cytomegalovirus after allogeneic hematopoietic stem cell transplantation: impact of host factors, drug

Assays for Quantitation of Plasma Cytomegalovirus DNAemia in Allogeneic Stem Cell Transplant Recipients. *J Clin Microbiol* Vol.49, No.8, pp. 2899-904, ISSN 1098- 660X


Caliendo, A. M.; Ingersoll, J.; Fox-Canale, A. M.; Pargman, S.; Bythwood, T.; Hayden, M. K.;

Caliendo, A. M.; Schuurman, R.; Yen-Lieberman, B.; Spector, S. A.; &ersen, J.; Manjiry, R.;

Caliendo, A. M.; Shahbazian, M. D.; Schaper, C.; Ingersoll, J.; Abdul-Ali, D.;

Caliendo, A. M.; St George, K.; Kao, S. Y.; Allega, J.; Tan, B. H.; LaFontaine, R.; Bui, L.; &

Cortez, K. J.; Fischer, S. H.; Fahle, G. A.; Calhoun, L. B.; Childs, R. W.; Barrett, A. J.; &

Deback, C.; Fillet, A. M.; Dhedin, N.; Barrou, B.; Varnous, S.; Najioullah, F.; Bricaire, F.; &

Drew, W. L. (2007). Laboratory diagnosis of cytomegalovirus infection and disease in

Emery, V. C.; & Griffiths, P. D. (2000). Prediction of cytomegalovirus load and resistance

Espy, M. J.; Uhl, J. R.; Sloan, L. M.; Buckwalter, S. P.; Jones, M. F.; Vetter, E. A.; Yao, J. D.;

Fahle, G. A.; & Fischer, S. H. (2000). Comparison of six commercial DNA extraction kits for

Flexman, J.; Kay, I.; Fonte, R.; Herrmann, R.; Gabbay, E.; & Palladino, S. (2001). Differences

*Microbiol Rev* Vol.19, No.1, pp. 165-256, ISSN 0893-8512

*J Clin Microbiol* Vol.45, No.6, pp. 1723-7, ISSN 0095-1137

No.4, pp. 1334-8, ISSN 0095-1137

1701-10, ISSN 1530-8561

2122-7, ISSN 0095-1137

Vol.40, No.3, pp. 173-9, ISSN 1386-6532

Vol.38, No.10, pp. 3860-3, ISSN 0095-1137

ISSN 0022-1899

0951-7375

8039-44, ISSN 0027-8424

660X

Assays for Quantitation of Plasma Cytomegalovirus DNAemia in Allogeneic Stem Cell Transplant Recipients. *J Clin Microbiol* Vol.49, No.8, pp. 2899-904, ISSN 1098-

Bremer, J. W.; & Lurain, N. S. (2007). Evaluation of real-time PCR laboratorydeveloped tests using analyte-specific reagents for cytomegalovirus quantification.

Crumpacker, C.; Lurain, N. S.; & Erice, A. (2001). Comparison of quantitative & qualitative PCR assays for cytomegalovirus DNA in plasma. *J Clin Microbiol* Vol.39,

Boonyaratanakornkit, J.; Pang, X. L.; Fox, J.; Preiksaitis, J.; & Schonbrunner, E. R. (2009). A commutable cytomegalovirus calibrator is required to improve the agreement of viral load values between laboratories. *Clin Chem* Vol.55, No.9, pp.

Rinaldo, C. R. (2000). Comparison of quantitative cytomegalovirus (CMV) PCR in plasma & CMV antigenemia assay: clinical utility of the prototype AMPLICOR CMV MONITOR test in transplant recipients. *J Clin Microbiol* Vol.38, No.6, pp.

Bennett, J. E. (2003). Clinical trial of quantitative real-time polymerase chain reaction for detection of cytomegalovirus in peripheral blood of allogeneic hematopoietic stem-cell transplant recipients. *J Infect Dis* Vol.188, No.7, pp. 967-72,

Agut, H. (2007). Monitoring of human cytomegalovirus infection in immunosuppressed patients using real-time PCR on whole blood. *J Clin Virol*

immunocompromised patients. *Curr Opin Infect Dis* Vol.20, No.4, pp. 408-11, ISSN

patterns after antiviral chemotherapy. *Proc Natl Acad Sci U S A* Vol.97, No.14, pp.

Wengenack, N. L.; Rosenblatt, J. E.; Cockerill, F. R. 3rd; & Smith, T. F. (2006). Realtime PCR in clinical microbiology: applications for routine laboratory testing. *Clin* 

recovery of cytomegalovirus DNA from spiked human specimens. *J Clin Microbiol*

between the quantitative antigenemia assay and the cobas amplicor monitor quantitative PCR assay for detecting CMV viraemia in bone marrow and solid organ transplant patients. *J Med Virol* Vol.64, No.3, pp. 275-82, ISSN 0146-6615


Detection of CMV Infection in Allogeneic SCT Recipients: The Multiple Assays 421

Kotton, C. N.; Kumar, D.; Caliendo, A. M.; Asberg, A.; Chou, S.; Snydman, D. R.; Allen, U.;

Ksouri, H.; Eljed, H.; Greco, A.; Lakhal, A.; Torjman, L.; Abdelkefi, A.; Ben Othmen, T.;

Leruez-Ville, M.; Ouachee, M.; Delarue, R.; Sauget, A. S.; Blanche, S.; Buzyn, A.; & Rouzioux,

Lilleri, D.; Baldanti, F.; Gatti, M.; Rovida, F.; Dossena, L.; De Grazia, S.; Torsellini, M.; &

Lilleri, D.; Lazzarotto, T.; Ghisetti, V.; Ravanini, P.; Capobianchi, M. R.; Baldanti, F.; &

Ljungman, P.; Griffiths, P.; & Paya, C. (2002). Definitions of cytomegalovirus infection and

Ljungman, P.; Perez-Bercoff, L.; Jonsson, J.; Avetisyan, G.; Sparrelid, E.; Aschan, J.; Barkholt,

Mackay, I. M.; Arden, K. E.; & Nitsche, A. (2002). Real-time PCR in virology. *Nucleic Acids* 

Machida, U.; Kami, M.; Fukui, T.; Kazuyama, Y.; Kinoshita, M.; Tanaka, Y.; K&a, Y.; Ogawa,

Martin-Davila, P.; Fortun, J.; Gutierrez, C.; Marti-Belda, P.; C&elas, A.; Honrubia, A.;

plasma. *J Clin Microbiol* Vol.41, No.5, pp. 2040-6, ISSN 0095-1137

ISSN 0041-1345

779-95, ISSN 1534-6080

pp. 1318-27, ISSN 0934-0874

Vol.183, No.3, pp. 377-82, ISSN 0022-1899

*Haematologica* Vol.91, No.1, pp. 78-83, ISSN 1592-8721

*Res* Vol.30, No.6, pp. 1292-305, ISSN 1362-4962

Vol.38, No.7, pp. 2536-42, ISSN 0095-1137

ISSN 0146-6615

6591

infection in kidney transplant recipients. *Transplant Proc* Vol.39, No.5, pp. 1458-60,

& Humar, A. (2010). International consensus guidelines on the management of cytomegalovirus in solid organ transplantation. *Transplantation* Vol.89, No.7, pp.

Ladeb, S.; Slim, A.; Zouari, B.; Abdeladhim, A.; & Ben Hassen, A. (2007). Analysis of cytomegalovirus (CMV) viremia using the pp65 antigenemia assay, the amplicor CMV test, and a semi-quantitative polymerase chain reaction test after allogeneic marrow transplantation. *Transpl Infect Dis* Vol.9, No.1, pp. 16-21, ISSN 1398-2273 Lehto, J. T.; Lemstrom, K.; Halme, M.; Lappalainen, M.; Lommi, J.; Sipponen, J.; Harjula, A.;

Tukiainen, P.; & Koskinen, P. K. (2005). A prospective study comparing cytomegalovirus antigenemia, DNAemia and RNAemia tests in guiding preemptive therapy in thoracic organ transplant recipients. *Transpl Int* Vol.18, No.12,

C. (2003). Monitoring cytomegalovirus infection in adult and pediatric bone marrow transplant recipients by a real-time PCR assay performed with blood

Gerna, G. (2004). Clinically-based determination of safe DNAemia cutoff levels for preemptive therapy or human cytomegalovirus infections in solid organ and hematopoietic stem cell transplant recipients. *J Med Virol* Vol.73, No.3, pp. 412-8,

Gerna, G. (2009). Multicenter quality control study for human cytomegalovirus DNAemia quantification. *New Microbiol* Vol.32, No.3, 245-53, ISSN 1121-7138 Limaye, A. P.; Huang, M. L.; Leisenring, W.; Stensl&, L.; Corey, L.; & Boeckh, M. (2001).

Cytomegalovirus (CMV) DNA load in plasma for the diagnosis of CMV disease before engraftment in hematopoietic stem-cell transplant recipients. *J Infect Dis*

disease in transplant recipients. *Clin Infect Dis* Vol.34, No.8, pp. 1094-7, ISSN 1537-

L.; Larsson, K.; Winiarski, J.; Yun, Z.; & Ringden, O. (2006). Risk factors for the development of cytomegalovirus disease after allogeneic stem cell transplantation.

S.; Honda, H.; Chiba, S.; Mitani, K.; Muto, Y.; Osumi, K.; Kimura, S.; & Hirai, H. (2000). Real-time automated PCR for early diagnosis and monitoring of cytomegalovirus infection after bone marrow transplantation. *J Clin Microbiol*

Barcena, R.; Martinez, A.; Puente, A.; de Vicente, E.; & Moreno, S. (2005). Analysis

therapy, and subclinical reactivation. *Blood* Vol.102, No.8, pp. 3060-7, ISSN 0006- 4971


Hanson, K. E.; Reller, L. B.; Kurtzberg, J.; Horwitz, M.; Long, G.; & Alex&er, B. D. (2007).

Harrington, S. M.; Buller, R. S.; Storch, G. A.; Li, L.; Fischer, S. H.; Murray, P. R.; & Gea-

Hebart, H.; & Einsele, H. (2004). Clinical aspects of CMV infection after stem cell

Herrmann, B.; Larsson, V. C.; Rubin, C. J.; Sund, F.; Eriksson, B. M.; Arvidson, J.; Yun, Z.;

cytomegalovirus. *J Clin Microbiol* Vol.42, No.5, pp. 1909-14, ISSN 0095-1137 Hong, K. M.; Najjar, H.; Hawley, M.; & Press, R. D. (2004). Quantitative real-time PCR with

Humar, A.; Gregson, D.; Caliendo, A. M.; McGeer, A.; Malkan, G.; Krajden, M.; Corey, P.;

Humar, A.; Kumar, D.; Boivin, G.; & Caliendo, A. M. (2002). Cytomegalovirus (CMV) virus

Ikewaki, J.; Ohtsuka, E.; Satou, T.; Kawano, R.; Ogata, M.; Kikuchi, H.; & Nasu, M. (2005).

Kaiser, L.; Perrin, L.; Chapuis, B.; Hadaya, K.; Kolarova, L.; Deffernez, C.; Huguet, S.; Helg,

DNA PCR assay. *J Clin Microbiol* Vol.40, No.11, pp. 4251-5, ISSN 0095-1137 Kalpoe, J. S.; Kroes, A. C.; de Jong, M. D.; Schinkel, J.; de Brouwer, C. S.; Beersma, M. F.; &

Kerschner, H.; Bauer, C.; Schlag, P.; Lee, S.; Goedel, S.; & Popow-Kraupp, T. (2011). Clinical

Kim, D. J.; Kim, S. J.; Park, J.; Choi, G. S.; Lee, S.; Kwon, C. D.; Ki, C.; & Joh, J. (2007). Real-

CMV disease. *J Infect Dis* Vol.186, No.6, pp. 829-33, ISSN 0022-1899

*Transplant* Vol.35, No.4, pp. 403-10, ISSN 0268-3369

recipients. *J Clin Microbiol* Vol.45, No.6, pp. 1972-3, ISSN 0095-1137

transplantation. *Hum Immunol* 65(5), pp. 432-6.

4971

ISSN 0009-9147

0095-1137

6, ISSN 1873-5967

pp. 1305-11, ISSN 0041-1337

therapy, and subclinical reactivation. *Blood* Vol.102, No.8, pp. 3060-7, ISSN 0006-

Comparison of the Digene Hybrid Capture System Cytomegalovirus (CMV) DNA (version 2.0), Roche CMV UL54 analyte-specific reagent, and QIAGEN RealArt CMV LightCycler PCR reagent tests using AcroMetrix OptiQuant CMV DNA quantification panels and specimens from allogeneic-stem-cell transplant

Banacloche, J. C. (2007). The effect of quantification standards used in real-time CMV PCR assays on guidelines for initiation of therapy in allogeneic stem cell transplant patients. *Bone Marrow Transplant* Vol.39, No.4, pp. 237-8, ISSN 0268-3369

Bondeson, K.; & Blomberg, J. (2004). Comparison of a duplex quantitative real-time PCR assay and the COBAS Amplicor CMV Monitor test for detection of

automated sample preparation for diagnosis & monitoring of cytomegalovirus infection in bone marrow transplant patients. *Clin Chem* Vol.50, No.5, pp. 846-56,

Greig, P.; Walmsley, S.; Levy, G.; & Mazzulli, T. (1999). Clinical utility of quantitative cytomegalovirus viral load determination for predicting cytomegalovirus disease in liver transplant recipients. *Transplantation* Vol.68, No.9,

load kinetics to predict recurrent disease in solid-organ transplant patients with

Real-time PCR assays based on distinct genomic regions for cytomegalovirus reactivation following hematopoietic stem cell transplantation. *Bone Marrow* 

C.; & Wunderli, W. (2002). Improved monitoring of cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation by an ultrasensitive plasma

Claas, E. C. (2004). 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* Vol.42, No.4, pp. 1498-504, ISSN

evaluation of a fully automated CMV PCR assay. *J Clin Virol* Vol.50, No.4, pp. 281-

time PCR assay compared with antigenemia assay for detecting cytomegalovirus

infection in kidney transplant recipients. *Transplant Proc* Vol.39, No.5, pp. 1458-60, ISSN 0041-1345


Detection of CMV Infection in Allogeneic SCT Recipients: The Multiple Assays 423

Preiser, W.; Brauninger, S.; Schwerdtfeger, R.; Ayliffe, U.; Garson, J. A.; Brink, N. S.; Franck,

Pumannova, M.; Roubalova, K.; Vitek, A.; & Sajdova, J. (2006). Comparison of quantitative

Raggam, R. B.; Bozic, M.; Salzer, H. J.; Hammerschmidt, S.; Homberg, C.; Ruzicka, K.; &

Razonable, R. R.; & Emery, V. C. (2004). Management of CMV infection and disease in

Ruell, J.; Barnes, C.; Mutton, K.; Foulkes, B.; Chang, J.; Cavet, J.; Guiver, M.; Menasce, L.;

Schaade, L.; Kockelkorn, P.; Ritter, K.; & Kleines, M. (2000). Detection of cytomegalovirus

Sia, I. G.; & Patel, R. (2000). New strategies for prevention & therapy of cytomegalovirus

Sia, I. G.; Wilson, J. A.; Espy, M. J.; Paya, C. V.; & Smith, T. F. (2000). Evaluation of the

Solano, C.; Munoz, I.; Gutierrez, A.; Farga, A.; Prosper, F.; Garcia-Conde, J.; Navarro, D.; &

Stocher, M.; Leb, V.; Bozic, M.; Kessler, H. H.; Halwachs-Baumann, G.; O.; Stekel, H.; & Berg,

Tanaka, N.; Kimura, H.; Iida, K.; Saito, Y.; Tsuge, I.; Yoshimi, A.; Matsuyama, T.; &

time PCR assay. *J Med Virol* Vol.60, No.4, pp. 455-62, ISSN 0146-6615

*Microbiol* Vol.39, No.11, pp. 3938-41, ISSN 0095-1137

PCR. *Med Microbiol Immunol* Vol.199, No.4, pp. 311-6, ISSN 1432-1831 Razonable, R. R.; Brown, R. A.; Wilson, J.; Groettum, C.; Kremers, W.; Espy, M.; Smith, T. F.;

disease. *Transplantation* Vol.73, No.6, pp. 968-73, ISSN 0041-1337

patients. *J Med Virol* Vol.76, No.3, pp. 367-72, ISSN 0146-6615

*Virol* Vol.20, No.1-2, pp. 59-70, ISSN 1386-6532

Vol.54, No.2, pp. 115-20, ISSN 0732-8893

7667

0268-3369

4006-9, ISSN 0095-1137

6, ISSN 0095-1137

1386-6532

No.1, 83-121, ISSN 0893-8512

in the monitoring of viral load during the antiviral treatment in renal transplant

S.; Doerr, H. W.; & Rabenau, H. F. (2001). Evaluation of diagnostic methods for the detection of cytomegalovirus in recipients of allogeneic stem cell transplants. *J Clin* 

competitive polymerase chain reaction-enzyme-linked immunosorbent assay with LightCycler-based polymerase chain reaction for measuring cytomegalovirus DNA in patients after hematopoietic stem cell transplantation. *Diagn Microbiol Infect Dis*

Kessler, H. H. (2010). Rapid quantitation of cytomegalovirus DNA in whole blood by a new molecular assay based on automated sample preparation & real-time

& Paya, C. V. (2002). The clinical use of various blood compartments for cytomegalovirus (CMV) DNA quantitation in transplant recipients with CMV

transplant patients. 27-29 February 2004. *Herpes* Vol.11, No.3, pp. 77-86, ISSN 0969-

Dougal, M.; & Chopra, R. (2007). Active CMV disease does not always correlate with viral load detection. *Bone Marrow Transplant* Vol.40, No.1, pp. 55-61, ISSN

DNA in human specimens by LightCycler PCR. *J Clin Microbiol* Vol.38, No.11, pp.

infection and disease in solid-organ transplant recipients. *Clin Microbiol Rev* Vol.13,

COBAS AMPLICOR CMV MONITOR test for detection of viral DNA in specimens taken from patients after liver transplantation. *J Clin Microbiol* Vol.38, No.2, pp. 600-

Gimeno, C. (2001). Qualitative plasma PCR assay (AMPLICOR CMV test) versus pp65 antigenemia assay for monitoring cytomegalovirus viremia & guiding preemptive ganciclovir therapy in allogeneic stem cell transplantation. *J Clin* 

J. (2003). Parallel detection of five human herpes virus DNAs by a set of real-time polymerase chain reactions in a single run. *J Clin Virol* Vol.26, No.1, pp. 85-93, ISSN

Morishima, T. (2000). Quantitative analysis of cytomegalovirus load using a real-

of quantitative PCR assay for CMV infection in liver transplant recipients: an intent to find the cut-off value. *J Clin Virol* Vol.33, pp. 138-144, ISSN 1386-6532


to find the cut-off value. *J Clin Virol* Vol.33, pp. 138-144, ISSN 1386-6532 Meijer, E.; Bol&, G. J.; & Verdonck, L. F. (2003). Prevention of cytomegalovirus disease in

Mengelle, C.; Mansuy, J. M.; Da Silva, I.; Davrinche, C.; & Izopet, J. (2011). Comparison of 2

Mengelle, C.; Pasquier, C.; Rostaing, L.; S&res-Saune, K.; Puel, J.; Berges, L.; Righi, L.;

Mhiri, L.; Kaabi, B.; Houimel, M.; Arrouji, Z.; & Slim, A. (2007). Comparison of pp65

Mori, T.; Okamoto, S.; Watanabe, R.; Yajima, T.; Iwao, Y.; Yamazaki, R.; Nakazato, T.; Sato,

Mori, T.; Sato, N.; Watanabe, R.; Okamoto, S.; & Ikeda, Y. (2000). Erythema exsudativum

Nicholson, V. A.; Whimbey, E.; Champlin, R.; Abi-Said, D.; Przepiorka, D.; Tarr&, J.; Chan,

Nitsche, A.; Steuer, N.; Schmidt, C. A.; L&t, O.; Ellerbrok, H.; Pauli, G.; & Siegert, W. (2000).

Nitsche, A.; Steuer, N.; Schmidt, C. A.; O.; & Siegert, W. (1999). Different real-time PCR

Ozdemir, E.; Saliba, R. M.; Champlin, R. E.; Couriel, D. R.; Giralt, S. A.; de Lima, M.; Khouri,

Pang, X. L.; Fox, J. D.; Fenton, J. M.; Miller, G. G.; Caliendo, A. M.; & Preiksaitis, J. K. (2009).

Piiparinen, H.; Helantera, I.; Lappalainen, M.; Suni, J.; Koskinen, P.; Gronhagen-Riska, C.; &

*Microbiol* Vol.38, No.7, pp. 2734-7, ISSN 0095-1137

*Clin Chem* Vol.45, No.11, pp. 1932-7, ISSN 0009-9147

Vol.9, No.2, pp. 258-68, ISSN 1600-6143

antigenemia. *J Med Virol* Vol.69, No.2, pp. 225-31, ISSN 0146-6615

*Bone Marrow Transplant* Vol.29, No.9, pp. 777-82, ISSN 0268-3369

647-57, ISSN 0893-8512

No.1, pp. 23-8, ISSN 0166-0934

ISSN 1879-0070

ISSN 0268-3369

ISSN 0268-3369

0268-3369

of quantitative PCR assay for CMV infection in liver transplant recipients: an intent

recipients of allogeneic stem cell transplants. *Clin Microbiol Rev* Vol.16, No.4, pp.

highly automated nucleic acid extraction systems for quantitation of human cytomegalovirus in whole blood. *Diagn Microbiol Infect Dis* Vol.69, No.2, pp. 161-6,

Bouquies, C.; & Izopet, J. (2003). Quantitation of human cytomegalovirus in recipients of solid organ transplants by real-time quantitative PCR and pp65

antigenemia, quantitative PCR and DNA hybrid capture for detection of cytomegalovirus in transplant recipients & AIDS patients. *J Virol Methods* Vol.143,

N.; Iguchi, T.; Nagayama, H.; Takayama, N.; Hibi, T.; & Ikeda, Y. (2002). Doseadjusted preemptive therapy for cytomegalovirus disease based on real-time polymerase chain reaction after allogeneic hematopoietic stem cell transplantation.

multiforme induced by granulocyte colony-stimulating factor in an allogeneic peripheral blood stem cell donor. *Bone Marrow Transplant* Vol.26, No.2, pp. 239-40,

K.; Bodey, G. P.; & Goodrich, J. M. (1997). Comparison of cytomegalovirus antigenemia & shell vial culture in allogeneic marrow transplantation recipients receiving ganciclovir prophylaxis. *Bone Marrow Transplant* Vol.19, No.1, pp. 37-41,

Detection of human cytomegalovirus DNA by real-time quantitative PCR. *J Clin* 

formats compared for the quantitative detection of human cytomegalovirus DNA.

I. F.; Hosing, C.; Kornblau, S. M.; Anderlini, P.; Shpall, E. J.; Qazilbash, M. H.; Molldrem, J. J.; Chemaly, R. F.; & Komanduri, K. V. (2007). Risk factors associated with late cytomegalovirus reactivation after allogeneic stem cell transplantation for hematological malignancies. *Bone Marrow Transplant* Vol.40, No.2, pp. 125-36, ISSN

Interlaboratory comparison of cytomegalovirus viral load assays. *Am J Transplant*

Lautenschlager, I. (2005). Quantitative PCR in the diagnosis of CMV infection and

in the monitoring of viral load during the antiviral treatment in renal transplant patients. *J Med Virol* Vol.76, No.3, pp. 367-72, ISSN 0146-6615


**19** 

*1,3USA 2Poland* 

**Bone Marrow Derived Pluripotent Stem Cells in** 

**Ischemic Heart Disease: Bridging the Gap** 

Ahmed Abdel-Latif1, Ewa Zuba-Surma2 and Mariusz Z. Ratajczak3

The prevalence of ischemic heart disease and acute myocardial infarction (AMI) has increased to alarming rates in the United States and the western world (Roger *et al.,* 2011). Patients who survive the initial AMI suffer ischemic cardiomyopathy (ICM) which is often complicated by high mortality and poor overall prognosis (Braunwald *et al.,* 2000; McMurray *et al.,* 2005). Despite significant advances in medical therapy and revascularization strategies, the prognosis of patients with AMI and ischemic cardiomyopathy remains dismal (Levy D *et al.,* 2002; Roger VL *et al.,* 2004). The last decade has demonstrated significant progress and rapid translation of myocardial regenerative therapies particularly those utilizing stem cells isolated from adult

Studies examining the potential therapeutic use of bone marrow (BM)-derived cells in myocardial regeneration have overshadowed the growing evidence of innate cardiac reparatory mechanisms. Several studies have demonstrated the capability of cardiomyocytes to replenish through poorly understood innate mechanisms. Follow up of cardiac transplantation patients have demonstrated continuous replenishment of cardiomyocytes by recipient's derived cells through poorly understood mechanisms (Quaini *et al.,* 2002). There is growing evidence that BM-derived cells are responsible, at least in part, for organ chimerism including cardiomyocyte chimerism (de Weger *et al.,* 2008; Deb *et al.,* 2003). Animal studies have confirmed this to be a dynamic process responding to significant injury such as myocardial infarction and peaks in the peri-infarct zone (Hsieh *et al.,* 2007). Although this process appears to be robust enough to achieve the renewal of approximately 50% of all cardiomyocytes in the normal life span, very little is known about its

**1. Introduction** 

tissues (Abdel-Latif *et al.,* 2007).

underpinnings (Bergmann *et al.,* 2009).

*University of Kentucky, and Lexington VA Medical Center, Lexington, KY* 

**Between Basic Research and** 

*1Gill Heart Institute and Division of Cardiovascular Medicine,* 

*2Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, 3Stem Cell Biology Institute, James Graham Brown Cancer Center,* 

**Clinical Applications** 

*University of Louisville, Louisville, KY,* 

