SYNTHESIS OF PCR-DERIVED, DIGOXIGENIN-LABELED DNA PROBES FOR IN-SITU DETECTION OF EPSTEIN-BARR EARLY RNAS IN EPSTEIN-BARR VIRUS-INFECTED CELLS

被引:37
作者
TSAI, ST
JIN, YT
WU, TC
机构
[1] NATL CHENG KUNG UNIV,COLL MED,DEPT PATHOL,TAINAN 70428,TAIWAN
[2] JOHNS HOPKINS UNIV,SCH MED,DEPT PATHOL,BALTIMORE,MD
关键词
DIGOXIGENIN; DNA PROBE; EPSTEIN-BARR EARLY RNA (EBER); POLYMERASE CHAIN REACTION;
D O I
10.1016/0166-0934(95)00030-X
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A PCR-derived digoxigenin-labeled DNA probe was used for for Epstein-Barr early RNA (EBER) in situ hybridization in formalin-fixed paraffin-embedded tissues. The results showed that the hybridization signal was morphologically distinct and the intensity of signal was comparable with those by RNA riboprobe. The advantages of using PCR-derived DNA probes for EBER in situ hybridization include: (1) the synthesis of digoxigenin-labeled DNA probes is easy and simple by PCR; (2) the labeled amplification product can be used as a probe without further purification; (3) DNA probes are potentially more stable than RNA probes; and (4) the preparation of DNA probes is relatively efficient and rapid. It is concluded that this technique is an ideal candidate for detection of EBER expression in clinical specimens.
引用
收藏
页码:67 / 74
页数:8
相关论文
共 26 条
[1]   GENERATION OF DIGOXIGENIN-LABELED DOUBLE-STRANDED AND SINGLE-STRANDED PROBES USING THE POLYMERASE CHAIN-REACTION [J].
AN, SF ;
FRANKLIN, D ;
FLEMING, KA .
MOLECULAR AND CELLULAR PROBES, 1992, 6 (03) :193-200
[2]   DNA-SEQUENCE AND EXPRESSION OF THE B95-8 EPSTEIN-BARR VIRUS GENOME [J].
BAER, R ;
BANKIER, AT ;
BIGGIN, MD ;
DEININGER, PL ;
FARRELL, PJ ;
GIBSON, TJ ;
HATFULL, G ;
HUDSON, GS ;
SATCHWELL, SC ;
SEGUIN, C ;
TUFFNELL, PS ;
BARRELL, BG .
NATURE, 1984, 310 (5974) :207-211
[3]   DETECTION OF MESSENGER-RNA AND LOW-ABUNDANCE HETERONUCLEAR RNA WITH SINGLE-STRANDED-DNA PROBES PRODUCED BY AMPLIFIED PRIMER EXTENSION LABELING [J].
BROOKS, PJ ;
KAPLITT, MG ;
KLEOPOULOS, SP ;
FUNABASHI, T ;
MOBBS, CV ;
PFAFF, DW .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1993, 41 (12) :1761-1766
[4]  
BROUSSET P, 1992, LAB INVEST, V67, P457
[5]  
EPSTEIN MA, 1964, LANCET, V1, P701
[6]   STRUCTURAL-ANALYSES OF EBER1 AND EBER2 RIBONUCLEOPROTEIN-PARTICLES PRESENT IN EPSTEIN-BARR VIRUS-INFECTED CELLS [J].
GLICKMAN, JN ;
HOWE, JG ;
STEITZ, JA .
JOURNAL OF VIROLOGY, 1988, 62 (03) :902-911
[7]   SYNTHESIS OF DIGOXIGENIN-LABELED DNA PROBE BY POLYMERASE CHAIN-REACTION - APPLICATION TO EPSTEIN-BARR-VIRUS AND CHLAMYDIA-TRACHOMATIS [J].
GRIFFAIS, R ;
ANDRE, PM ;
THIBON, M .
RESEARCH IN VIROLOGY, 1990, 141 (03) :331-335
[8]   SYNTHESIS OF PCR-DERIVED, SINGLE-STRANDED-DNA PROBES SUITABLE FOR IN-SITU HYBRIDIZATION [J].
HANNON, K ;
JOHNSTONE, E ;
CRAFT, LS ;
LITTLE, SP ;
SMITH, CK ;
HEIMAN, ML ;
SANTERRE, RF .
ANALYTICAL BIOCHEMISTRY, 1993, 212 (02) :421-427
[9]   RELATION OF BURKITTS TUMOR-ASSOCIATED HERPES-TYPE VIRUS TO INFECTIOUS MONONUCLEOSIS [J].
HENLE, G ;
HENLE, W ;
DIEHL, V .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1968, 59 (01) :94-&
[10]   LOCALIZATION OF EPSTEIN-BARR VIRUS-ENCODED SMALL RNAS BY INSITU HYBRIDIZATION [J].
HOWE, JG ;
STEITZ, JA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (23) :9006-9010