Standardization of the Human Cytomegalovirus Antigenemia Assay by Means of In Vitro-Generated pp65-Positive Peripheral Blood Polymorphonuclear Leukocytes

Journal of Clinical Microbiology - Tập 36 Số 12 - Trang 3585-3589 - 1998
Giuseppe Gerna1, Elena Percivalle1, Maria Torsellini1, Maria Grazia Revello1
1Servizio di Virologia, Istituto di Ricovero e Cura a Carattere Scientifico Policlinico S. Matteo, 27100 Pavia, Italy

Tóm tắt

ABSTRACT We generated in vitro human cytomegalovirus (HCMV) pp65-positive polymorphonuclear leukocytes (PMN) resembling those detected in vivo, following cocultivation of PMN from healthy donors and wild-type HCMV-infected endothelial cells or fibroblasts. After purification, PMN are suitable for preparation of cytospots which can be used for the antigenemia assay. Cytospin preparations containing a predetermined number of in vitro-generated pp65-positive PMN were used to test some of the major parameters involved in performing the antigenemia assay. The results showed or confirmed that (i) formalin fixation followed by permeabilization is the best fixation procedure developed to date, (ii) the test performance levels provided by different pools of pp65-specific monoclonal antibodies may be significantly different, and (iii) long-term storage (for an unlimited time) is best achieved by keeping fixed slides at −80°C, whereas short-term storage (for up to 1 month) is best achieved by keeping unfixed slides at room temperature. This finding signifies that slides can be shipped all over the world at room temperature. In conclusion, the newly developed procedure for in vitro generation of pp65-positive PMN will provide the basis for standardization of the HCMV antigenemia assay and development of quality control programs.

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Tài liệu tham khảo

10.1128/jcm.32.3.832-834.1994

10.1128/jcm.35.7.1819-1821.1997

10.1128/jcm.30.5.1232-1237.1992

10.1099/0022-1317-73-11-2923

10.1128/jcm.34.5.1337-1339.1996

10.1128/jcm.33.1.257-259.1995

10.1128/jcm.32.4.1119-1120.1994

10.1128/jcm.33.6.1646-1649.1995

10.1172/JCI1633

10.1128/jcm.35.3.741-744.1997

10.1016/0092-8674(94)90337-9

The T. H. van der Berg A. P. Harmsen M. C. van der Bij W. van Son W. J. The cytomegalovirus antigenemia assay: a plea for standardization.Scand. J. Infect. Dis. Suppl.9919952529

10.1128/jcm.26.12.2531-2535.1988

10.1002/jmv.1890250208

10.1128/jcm.29.12.2909-2910.1991

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Thông tin xuất bản

Journal of Clinical Microbiology

Tập 36 Số 12

3585-3589

Thông tin tác giả