Zbornik - Društvo genetičara Srbije

V-Pos-12 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 215
Subotica, 30. novembar - 4. decembar 2004.
PCR DETEKCIJA GENOMA RAZLIÈITIH MIKROORGANIZAMA U
SINOVIJI BOLESNIKA SA REITER-OVIM SINDROMOM TRETIRANIH
AZITROMICINOM
N. Streliæ 1 , Lj. Pavlica 2 i Z. Magiæ 1
1
Institut za medicinska istraivanja, Beograd
2
Klinika za reumatologiju, Vojnomedicinska akademija, Beograd
Bakterijska infekcija zgloba je ozbiljan problem èiji ishod zavisi od pravilnog tretmana.
Mali broj intra-artikularnih bakterija se moe detektovati ultraosetljivim molekularnim
tehnikama, baziranim na lanèanoj reakciji polimeraze (PCR). Brza identifikacija
bakterijske infekcije je neophodna za odgovarajuæi tretman inficiranih pacijenata.
Cilj ovog istraivanja je bio detekcija bakterijske DNK u tkivu sinovije, sinovijskoj
teènosti i perifernoj krvi bolesnika sa Reiter-ovim sindromom tretiranih azitromicinom.
Tkivo sinovije, sinovijska teènost i mononukleari periferne krvi su korišæeni kao izvori
DNK u PCR amplifikaciji bakterijskog 16 S rRNK gena. PCR prajmeri za specifiènu
amplifikaciju DNK fragmenta 16S rRNK gena razlièitih bakterija (Chlamydia
trachomatis, C. pneumoniae, Mycoplasma hominis, Ureaplasma urealyticum) su bili
visoko specifièni. Uzorci su dobijeni od 20 bolesnika sa Reiter-ovim sindromom. DNK
je izolovana fenol-hloroform ekstrakcijom i posle PCR amplifikacije bakterijska DNK je
detektovana na 10% PAGE.
Bakterijska DNK je detektovana u tri uzorka sinovijske teènosti (15%), 11 uzoraka tkiva
sinovije (55%) i 17 uzoraka periferne krvi (80%). Nakon tretmana azitromicinom,
bakterijska DNK je detektovana u tri uzorka sinovijske teènosti (15%), 7 uzoraka tkiva
sinovije (35%) i 10 uzoraka periferne krvi (50%).
Najbolji rezultat terapije azitromicinom je dobijen u krvi posebno za Mycoplasma
hominis.
PCR DETECTION OF GENOME OF DIFFERENT MICROORGANISMS IN
SYNOVIA OF PATIENTS WITH REITER’S SYNDROME TREATED WITH
AZITHROMYCIN
Bacterial joint infection is a serious problem, the outcome of which depends on appropriate
treatment. A small number of intra-articular bacteria can only be detected by using
ultrasensitive molecular techniques, mainly those based on the polymerase chain reaction
(PCR). The rapid identification of bacterial infection is essential for the proper treatment
of infected patients.
The aim of this study was to detect bacterial DNA in synovial tissue, synovial fluid and
peripheral blood of patients with Reiter's syndrome treated with azithromycin.
Synovial tissue, synovial fluid and peripheral blood mononuclear cells were used as the
source of DNA for PCR amplification of bacterial 16 S rRNA gene. PCR primers for the
specific amplification of DNA fragment of the 16 S rRNA gene of different bacteria
(Chlamydia trachomatis, C. pneumoniae, Mycoplasma hominis, Ureaplasma urealyticum)
were highly specific. Samples were obtained from 20 patients with Reiter’s syndrome.
DNA was isolated by phenol/chloroform extraction and after PCR amplification
bacterial DNA was detected by 10% PAGE.
Bacterial DNA was detected in 3 samples of the synovial fluid (15%), 11 samples of the
synovial tissue (55%), and 17 samples of peripheral blood (80%). After the treatment
with azithromycin, bacterial DNA was detected in 3 samples of the synovial fluid (15%),
7 samples of the synovial tissue (35%), and 10 samples of peripheral blood (50%).
The best result with azithromycin therapy was obtained in blood especially for
Mycoplasma hominis.