Naslov
Development of a new method for identification of uropathogenic E. coli (P-type)

Autori
Dumić Jerka ; Lauc Gordan ; Lee Yuan C. ; Flögel, Mirna

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
CCOMLIS1, 1st Croatian Congress on Molecular Life Sciences, Opatija, Croatia, June 09-14, 2002. Book of Abstracts / Dumić, Jerka ; Kućan, Željko ; Fögel, Mirna - Zagreb : Farmaceutsko-biokemijski fakultet Sveučilišta u Zagrebu, 2002

Skup
1st CCOMLIS, 1st Croatian Congress on Molecular Life Sciences

Mjesto i datum
Opatija, Hrvatska, 09.06.2002. - 13.06.2002

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
uropathogenic E. coli; analysis; method

Sažetak
E. coli is by far the most common cause of urinary tract infections. Standard methods for identification of uropathogenic E. coli (UPEC) in urine are semi-quantitative and usually take 2-3 days, so development of a more rapid, molecular approach to detect UPEC may lead to earlier diagnosis and improve management of urinary tract infections. Attachment of UPEC to host cell receptors is mediated by interaction of the bacterial lectin (adhesin) localized at the top of fimbriae and specific carbohydrate moiety of the host membrane glycoconjugates. The most common type of UPEC causing pyelonephritis expresses type P adhesin (UPEC-P) that specifically recognizes P1 antigen (Galα (1-4)Gal moiety – galabiose). Pigeon glycoproteins are rich in galabiose sequences at the terminal positions of N-glycans that makes them potent inhibitors of UPEC-P adhesion. That fact, together with basic principles of “ sandwich ELISA” -method, served us as a basis for developing a new method for UPEC-P detection. To achieve specific binding of UPEC-P, microtitar plates were coated with purified pigeon ovalbumin. Next step comprised application of ovalbumin N-glycans conjugated with digoxin assuring in that way specific labeling of bound bacteria. According to our knowledge this is the first case of glycans labeling with digoxin as well as their application. Visualization was achieved with antibodies against digoxigenin (that specifically recognize digoxin, too) conjugated with alkaline phosphatase and p-nitrophenyl phosphate as a substrate. Our preliminary study indicates the feasibility of using specific glycoproteins and glycans labeled with digoxin for development of a rapid, species-specific molecular approach to detect uropathogenic E. coli. The sensitivity of such approach may be further utilized for detection of other bacterial species, measuring of bacterial virulence as well as antibiotic sensitivity.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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