| Sažetak | CILJ: Cilj ovog završnog rada bio je predstaviti molekularne metode dijagnostike infekcija središnjeg živčanog sustava te ih usporediti s standardnim mikrobiološkim metodama. Također, rad prikazuje molekularne metode u sklopu sindromskog pristupa dijagnostici te daje presjek infekcija središnjeg živčanog sustava. MATERIJALI I METODE: Završni rad pisan je pomoću stručne literature dobivene pretraživanjem znanstvenih baza na internetu (PubMed, Cochrane Database of Systematic Reviews, Hrčak, CROSBI), stručnih udžbenika te priručnika prikladnih tema. Literatura je korištena iz raznih grana medicine i laboratorijske dijagnostike. Pretraga literature dijelom je vođena PRISMA smjernicama. REZULTATI: Molekularna dijagnostika u vidu dijagnostike infekcija središnjeg živčanog sustava sve više izbacuje standardne mikrobiološke metode iz svakodnevne prakse. Svojom visokom osjetljivošću i specifičnošću puno brže detektira patogene, što smanjuje troškove, olakšava liječenje te skraćuje boravak u bolnici. Shodno tome, molekularna dijagnostika danas postaje novi „zlatni standard“ za dijagnostiku različitih zaraznih bolesti, a između ostalog i infekcija središnjeg živčanog sustava. Metode koje se primjenjuju u molekularnoj dijagnostici jesu: tehnike hibridizacije, tehnike amplifikacije signala, tehnike amplifikacija nukleinskih kiselina (kao što je, primjerice, metoda lančane reakcije polimerazom ili PCR) te tehnike sekvenciranja dijelova genoma ili čitavog genoma uzročnika. Dominantno se u dijagnostici infekcija središnjeg živčanog sustava koristi tehnika lančane reakcije polimerazom u stvarnom vremenu (real-time PCR). ZAKLJUČAK: Standardni PCR, multiplex PCR te metagenomske metode dijagnostike svojom brzinom, specifičnošću i osjetljivošću pomažu ne samo u detekciji patogena već ih možemo koristiti i za praćenje terapije, genotipizaciju, za analizu rezistentnosti mikroorganizama pa čak i za indirektno praćenje kinetike replikacije. Svojim napretkom postaju „zlatni standard“ mikrobiološke dijagnostike te zamjenjuju standardne metode koje su sporije te kod kojih postoji veća šansa za kontaminaciju. Otvorenom suradnjom kliničara i prvostupnika medicinsko-laboratorijske dijagnostike postiže se brža i efikasnija dijagnostika infekcija središnjeg živčanog sustava uz pomoć molekularnih metoda. KLJUČNE RIJEČI: središnji živčani sustav, infekcija, meningitis, encefalitis, mikrobiologija, dijagnostika, molekularna dijagnostka 14. SUMMARY 23 14. SUMMARY Molecular diagnostic approaches for central nervous system infections AIM: The aim of this thesis was to present molecular methods for diagnosing central nervous system infections and to compare them with standard microbiological methods. Also, the thesis presents molecular methods as an inherent part of a syndromic approach to diagnose these infections, and gives a review of central nervous system infections. MATERIALS AND METHODS: The thesis has used studies obtained by searching scientific databases on the Internet (PubMed, Cochrane Database of Systematic Reviews, Hrčak, CROSBI), professional textbooks and manuals on relevant topics. The literature that has been used covers various branches of medicine and laboratory diagnostics. PRISMA guidelines were partly consulted for the development of search strategy. RESULTS: Molecular diagnostics of the central nervous system infections is increasingly replacing standard microbiological methods from the quotidian practice. With their high sensitivity and specificity, pathogen detection is much faster, which reduces costs, facilitates treatment and shortens hospital stay. Consequently, molecular diagnostics is becoming the new “gold standard” for the diagnosis of various infectious diseases – including central nervous system infections. Methods used in such molecular diagnostic approaches are: hybridization techniques, signal amplification techniques, nucleic acid amplification techniques (such as polymerase chain reaction or PCR) and sequencing techniques of parts of the genome or the entire pathogen genome. The technique of real-time PCR polymerase chain reaction is predominantly used in the diagnosis of central nervous system infections. CONCLUSION: Due to their speed, specificity and sensitivity, standard PCR, multiplex PCR and metagenomic diagnostic methods aid not only in pathogen detection, but can also be used to monitor therapy, for genotyping purposes, to analyze resistance of microorganisms and even to indirectly monitor replication kinetics. The technological progress makes them a “gold standard” of microbiological diagnostics and replaces standard methods that are slower and have a higher chance of contamination. Through the open cooperation of clinicians and bachelors of medical-laboratory diagnostics, faster and more efficient diagnosis of infections of the central nervous system can be achieved with the use of molecular methods. |
| Sažetak (engleski) | AIM: The aim of this thesis was to present molecular methods for diagnosing central nervous system infections and to compare them with standard microbiological methods. Also, the thesis presents molecular methods as an inherent part of a syndromic approach to diagnose these infections, and gives a review of central nervous system infections. MATERIALS AND METHODS: The thesis has used studies obtained by searching scientific databases on the Internet (PubMed, Cochrane Database of Systematic Reviews, Hrčak, CROSBI), professional textbooks and manuals on relevant topics. The literature that has been used covers various branches of medicine and laboratory diagnostics. PRISMA guidelines were partly consulted for the development of search strategy. RESULTS: Molecular diagnostics of the central nervous system infections is increasingly replacing standard microbiological methods from the quotidian practice. With their high sensitivity and specificity, pathogen detection is much faster, which reduces costs, facilitates treatment and shortens hospital stay. Consequently, molecular diagnostics is becoming the new “gold standard” for the diagnosis of various infectious diseases – including central nervous system infections. Methods used in such molecular diagnostic approaches are: hybridization techniques, signal amplification techniques, nucleic acid amplification techniques (such as polymerase chain reaction or PCR) and sequencing techniques of parts of the genome or the entire pathogen genome. The technique of real-time PCR polymerase chain reaction is predominantly used in the diagnosis of central nervous system infections. CONCLUSION: Due to their speed, specificity and sensitivity, standard PCR, multiplex PCR and metagenomic diagnostic methods aid not only in pathogen detection, but can also be used to monitor therapy, for genotyping purposes, to analyze resistance of microorganisms and even to indirectly monitor replication kinetics. The technological progress makes them a “gold standard” of microbiological diagnostics and replaces standard methods that are slower and have a higher chance of contamination. Through the open cooperation of clinicians and bachelors of medical-laboratory diagnostics, faster and more efficient diagnosis of infections of the central nervous system can be achieved with the use of molecular methods. |
