Validnost različitih laboratorijskih metoda za dijagnozu onihomikoza i identifikacija faktora rizika

Abstract

Uvod: Onihomikoza predstavlja gljivičnu infekciju jednog ili više noktiju koja je izazvana bilo kojom vrstom gljiva: dermatofitima, kvasnicama ili nedermatofitnim plesnima. Oboljenje može da zahvati bilo koji deo nokta, uključujući matriks, nokatno ležište i/ili nokatnu ploču. Procene su da oko 5% svetske populacije ima neki oblik onihomikoze i zbog toga ova bolest predstavlja značajan zdravstveni problem. Novije studije ukazuju da je u zemljama zapadne i centralne Evrope prevalencija onihomikoza 3 – 22%. Kada je u pitanju Srbija, ne postoje podaci o učestalosti onihomikoza niti o najčešćim uzročnicima ovog oboljenja kod naših pacijenata. Ciljevi: Određivanje učestalosti javljanja onihomikoza, kao i najčešćih uzročnika onihomikoza kod pacijenata sa kliničkom sumnjom na onihomikozu, analiziranje validnosti različitih laboratorijskih metoda za dijagnozu onihomikoza, ispitivanje uticaja primene tripsina na bolju identifikaciju uzročnika onihomikoza, identifikovanje faktora koji doprinose nastajanju onihomikoza. Materijal i metode: Studija slučajeva obuhvatila je 374 pacijenata sa kliničkom sumnjom na onihomikozu iz šest ustanova: Institut za Mikrobiologiju i imunologiju, Medicinski fakultet Univerziteta u Beogradu; Zavod za javno zdravlje, Pančevo; Opšta bolnica „Dr Đorđe Joanović“, Zrenjanin; Opšta bolnica, Užice; Zavod za javno zdravlje „Timok“, Zaječar; Zavod za javno zdravlje, Kraljevo. Podaci od pacijenata su prikupljeni odgovarajućim anketnim upitnikom. Od svakog pacijenta uzet je uzorak nokta u zavisnosti od kliničkog tipa onihomikoze i poslat u Institut za mikrobiologiju i imunologiju, Medicinskog fakulteta Univerziteta u Beogradu gde je vršena dalja obrada materijala. U cilju identifikovanja uzročnog faktora onihomikoze za svaki uzorak nokta urađeni su sledeći laboratorijski testovi, sa i bez prethodnog tretiranja tripsinom: direktan mikroskopski preparat sa KOH, direktan mikroskopski preparat sa Blankophor-om, kultivisanje na neselektivnim mikološkim podlogama SDA i RSDA, kao i selektivnoj podlozi za dermatofite DTM. Svaki uzorak nokta je testiran i pomoću Dermatophyte PCR kita kojim je moguće identifikovati dermatofite uopšteno (panderm marker), kao i specifično Trichophyton rubrum. Kriterijum za postavljanje dijagnoze onihomikoze podrazumevao je postojanje jasne kliničke sumnje na onihomikozu uz pozitivnost bilo kog laboratorijskog testa...

Background: Onychomycosis represents fungal infection of one or more nails caused by any type of fungi: dermatophytes, yeasts or nondermatophyte molds. The disease can affect any part of the nail, including matrix, nail bed and/or nail plate. It is estimated that about 5% of the world's population has some form of onychomycosis which makes it a significant health problem. Recent studies indicate that in the countries of Western and Central Europe, the prevalence of onychomycosis is 3-22%. Regarding Serbia, there are no data on frequency of onychomycosis nor about the most common causes of this disease in our patients. Aims: Determination of frequency of onychomycosis, as well as the most common causes of onychomycosis in patients with clinical suspicion of onychomycosis, analyzing the validity of the different laboratory methods in the diagnosis of onychomycosis, investigation of the impact of application of trypsin in better identification of causativ agents of onychomycosis, identification of risk factors associated with onychomycosis. Material and methods: The case study included 374 patients with clinical suspicion of onychomycosis from six institutions: Institute of Microbiology and Immunology, Faculty of Medicine, University of Belgrade; Department of Public Health, Pančevo; General Hospital 'Dr Đorđe Joanović", Zrenjanin; General Hospital Užice; Public Health Institute "Timok", Zaječar; Department of Public Health, Kraljevo. Data from patients was collected by appropriate questionnaire. From each patient a nail sample was taken according to clinical type of onychomycosis and was sent to the Institute of Microbiology and Immunology, Faculty of Medicine, University of Belgrade, where further processing of the material was done. In order to identify the causative agent of onychomycosis each nail sample underwent the following laboratory tests, with and without previous treatment with trypsin: direct microscopic examination with KOH, direct microscopic examination with Blankophor, culturing on non-selective mycologic media SDA and RSDA, and on selective medium for dermatophytes DTM. Each nail sample was also tested by Dermatophyte PCR Kit which can identify dermatophytes in general (panderm marker), as well as the specific identification of Trichophyton rubrum. The criteria for the diagnosis of onychomycosis implied the existence of clear clinical suspicion of onychomycosis with positivity of any laboratory test. Based on laboratory diagnosis, subjects were divided into two groups: patients with a laboratory confirmed diagnosis of onychomycosis and subjects without the disease, in order to identify risk factors for onychomycosis. Clinical SCIO index was calculated for each patient...