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Validacija multiplega testa PCR v realnem času za dokaz okužb dihal z atipičnimi bakterijami
ID Brodarič, Jakob (Author), ID Keše, Darja (Mentor) More about this mentor... This link opens in a new window

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Abstract
Atipične bakterije, kot so Chlamydia pneumoniae, Mycoplasma pneumoniae in Legionella pneumophila, so pomembni povzročitelji okužb dihal. Okužbe lahko potekajo brezsimptomno, z blagimi simptomi ali pa se razvijejo v težjo obliko pljučnice, ki zahteva bolnišnično oskrbo. Ker so omenjeni patogeni odporni na betalaktamske antibiotike, je za zdravljenje bolnikov pomembno, da jih kot povzročitelje okužbe odkrijemo dovolj zgodaj. Zaradi posebnosti v njihovi celični zgradbi in pogojih razmnoževanja jih težko izoliramo iz kliničnih vzorcev; metoda osamitve je zahtevna, dolgotrajna in ima nizko občutljivost. Prav tako so zamudni serološki testi, saj potrebujemo parne serume za potrditev akutne okužbe. Diagnostika se zato izvaja večinoma z molekularnimi testi, kot je multipli test PCR, ki omogoča hkratno prepoznavanje okužb s katerokoli atipično bakterijo. Namen naloge je bil ovrednotiti novejše diagnostične multiple teste PCR proizvajalcev TIB MolBiol, Seegene, Fast Track in AusDiagnostic, za katere smo predvidevali, da bodo imeli višjo občutljivost od testa, ki je trenutno v uporabi, in bodo enostavnejši za izvedbo. Rezultati niso potrdili višje občutljivosti od trenutno uporabljanega testa proizvajalca Argene, so pa pokazali, da so omenjeni novejši testi enostavnejši za uporabo.

Language:Slovenian
Keywords:medicinska mikrobiologija, atipične bakterije, Mycoplasma pneumoniae, Chlamydia pneumoniae, Legionella pneumophila, diagnostične metode, molekularne tehnike, PCR v realnem času, validacija
Work type:Master's thesis/paper (mb22)
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2021
Publisher:[J. Brodarič]
Place:Ljubljana
UDC:579.61:616.2-078:577.2.083
COBISS.SI-ID:61739523 This link opens in a new window
Publication date in RUL:30.04.2021
Views:341
Downloads:79
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Secondary language

Language:English
Title:Validation of a multiplex real-time PCR for detection of atypical bacteria causing respiratory infections
Abstract:
Atypical bacteria, such as Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella pneumophila, are major causes of respiratory infections. Infections can be asymptomatic, with mild symptoms or can further develop into a severe form of pneumonia requiring hospital care. Since the afore mentioned pathogens are resistant to beta-lactam antibiotics, it is of great importance for patients’ treatment that they be identified early enough as infection causes. Due to their specific cellular structure and multiplication conditions, it is difficult to separate them from the clinical samples; to be more precise, the isolation method is challenging, time-consuming, and has a low sensitivity rate. Serologic tests are also lengthy, for in order to confirm the acute infection, steam sera are needed. Consequently, the diagnostics are mostly carried out with molecular tests such as multiplex PCR test which enables the simultaneous detection of infections with whichever atypical bacterium. The objective of this thesis is to evaluate more recent diagnostic multiplex PCR tests by TIB MolBiol, Seegene, Fast Track and AusDiagnostic. Our predictions were that they would have a higher sensitivity detection than the test that is currently in use, in addition to being easier to perform. The results have shown that not all tests are more sensitive than the Argene test and confirmed that they really are more user-friendly.

Keywords:medical microbiology, atypical bacteria, Mycoplasma pneumoniae, Chlamydia pneumoniae, Legionella pneumophila, diagnostic methods, molecular techniques, real-time PCR

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