Pregled bibliografske jedinice broj: 55071
Nonradioactive digoxigenin DNA labeling and imunological detection of HSV PCR products
Nonradioactive digoxigenin DNA labeling and imunological detection of HSV PCR products // 1st Croatian congress on infectious diseases with international participation / Croatian medical association (ur.).
Dubrovnik: Croatian society of infectology, 1998. (poster, domaća recenzija, sažetak, znanstveni)
CROSBI ID: 55071 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Nonradioactive digoxigenin DNA labeling and imunological detection of HSV PCR products
Autori
Merle-Broketa, Martina ; Vince, Adriana ; Draženović, Vladimir ; Mlinarić- Galinović, Gordana
Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni
Izvornik
1st Croatian congress on infectious diseases with international participation
/ Croatian medical association - Dubrovnik : Croatian society of infectology, 1998
Skup
1st Croatian congress on infectious diseases with international participation
Mjesto i datum
Dubrovnik, Hrvatska, 01.10.1998. - 03.10.1998
Vrsta sudjelovanja
Poster
Vrsta recenzije
Domaća recenzija
Ključne riječi
nonradioactive digoxigenin DNA labeling;HSV PCR products
Sažetak
A nonradioactive molecular assay for detection of Herpes Simplex Virus (HSV) amplification products was evaluated. After phenol-chloroform DNA extraction from green monkey kidney (GMK) cell cultures infected with HSV-1, specific DNA sequence was amplified by polymerase chain reaction (PCR), using specific oligonucleotide primers (OPERON, Operon Technologies, Inc., Alameda, CA). Amplification products were visualized by agarose gel electrophoresis, and if present, specific DNA HSV probe (OPERON, Operon Technologies, Inc., Alameda, CA) was labeled. DNA probe is labeled by random primed incorporation of digoxigenin-labeled deoxyuridine triphosphate (DIG DNA labeling and detection kit, Boehringer Mannheim GmbH). The dUTP is linked via a spacer-arm to the steroid hapten digoxigenin (DIG-dUTP). Southern transfer to positively charged nylon membrane (Boehringer Mannheim Nylon Membran) or Dot blot were performed. DIG-labeled DNA is used according to standard hybridization protocol. After hybridization to the target DNA hybrids were detected by enzyme-linked immunoassay using an antibody-conjugate (antidigoxigenin-alkaline phosphatase conjugate) and subsequent enzyme-catalyzed color reaction with 5-bromo-4-chloro-3-indolyl phosphate (X-phosphate) and nitro blue tetrazolium salt (NBT).
Izvorni jezik
Engleski
Znanstvena područja
Kliničke medicinske znanosti
