Pretreatment with restriction enzyme or bovine serum albumin for effective PCR amplification of Epstein-Barr virus DNA in DNA extracted from paraffin-embedded gastric carcinoma tissue.
An association between Epstein-Barr virus (EBV) and gastric carcinoma has been studied through the EBV genome present in the carcinoma cells. Recently, we found that EBV DNA in paraffin-embedded gastric carcinoma tissue was detected effectively by PCR after pretreatment of the extracted DNA with a restriction enzyme, BamHI or EcoRI. Here, we show that the PCR amplification was also enhanced by pretreatment of the DNA with other restriction enzymes or with bovine serum albumin and several other proteins. Treatment with these proteins may remove a PCR inhibitor(s) in the DNA samples extracted from the paraffin blocks.
Departments of Biosignaling, Faculty of Medicine, Tottori University, Yonago 683, Japan.
Y
Y
Satoh
N
N
Takasaka
Y
Y
Hoshikawa
M
M
Osaki
S
S
Ohfuji
H
H
Ito
N
N
Kaibara
T
T
Kurata
T
T
Sairenji
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification*; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI; D020031:Epstein-Barr Virus Infections / Q000150:complications; D005069:Evaluation Studies as Topic; D005914:Globins / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000302:isolation & purification* / Q000472:pathogenicity; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012710:Serum Albumin, Bovine; D013274:Stomach Neoplasms / Q000209:etiology / Q000821:virology*; D014773:Virology / Q000379:methods
D017931:DNA Primers; D004279:DNA, Viral; D012710:Serum Albumin, Bovine; D005914:Globins; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI