Protokol usposabljanja laboratorijskih delavcev za ocenjevanje dvojnega p16/Ki-67 imunocitokemičnega barvanja v brisih materničnega vratu

Rozina, Hermina (author), Kloboves Prevodnik, Veronika (mentor)

Presejalni test p16/Ki-67 je dvojno imunocitokemično barvanje (ICK) in eden od obetavnih testov, ki bi jih lahko v prihodnosti uporabljali za odkrivanje predrakavih sprememb in raka na materničnem vratu. Za njegovo varno uporabo so na Onkološkem inštitutu Ljubljana razvili program usposabljanja laboratorijskih delavcev na tem področju. V raziskavo smo vključili dva učenca brez znanja na področju citopatologije in starejšega presejalca (SP). Raziskava je bila sestavljena iz dveh delov. V prvem delu sta bila učenca deležna kratkega izobraževalnega programa o osnovah normalne ginekološke citopatologije in načinu ocenjevanja dvojnega p16/Ki-67 ICK barvanja. Pregledala sta tudi 118 preparatov, pobarvanih z dvojnim p16/Ki-67 ICK barvanjem, brez možnosti sprotnega posvetovanja s SP. V drugem delu sta s SP na diskusijskem mikroskopu naprej prediskutirala preparate z neskladnimi rezultati iz prvega ocenjevanja in nato pregledala še 383 preparatov. Med drugim ocenjevanjem sta se lahko sproti posvetovala s SP. Primerjali smo skladnost med ocenjevalci in referenco ter izračunali občutljivost in specifičnost dvojnega p16/Ki-67 barvanja za detekcijo CIN2+. Spremljali smo tudi čas pregledovanja posameznega preparata pri učencih. S statističnimi metodami smo dokazali, da se je v začetnem ocenjevanju referenci najbolj približal SP, v dodatnem pa učenec 2 (U2). Občutljivost se je po dodatnem ocenjevanju pri obeh učencih znižala zaradi povečane specifičnosti. V primerjavi z referenco so po dodatnem ocenjevanju še vedno vsi ocenjevalci imeli nižjo občutljivost in specifičnost. Boljši čas pregledovanja pri zadnjih 100 preparatih je dosegel U2. Po končanem testiranju je bil U2 usposobljen za samostojno pregledovanje preparatov z dvojnim p16/Ki-67 ICK barvanjem, medtem ko U1 ni bil primeren za samostojno delo, saj bi se moral še dodatno izobraževati in delati pod nadzorom SP.

• Language: Slovenian
• Keywords: p16, Ki-67, rak materničnega vratu, imunocitokemija, humani papiloma virus
• Work type: Master's thesis/paper
• Organization: BF - Biotechnical Faculty
• Year: 2019
• PID: 20.500.12556/RUL-111198
• COBISS.SI-ID: 5301839
• Publication date in RUL: 26.09.2019

Secondary language

• Language: English
• Title: Assessment of in-house training protocol for the evaluation of dual p16/Ki-67 immunocytochemical staining in cervical smears

Abstract

p16/Ki-67 dual immunocytochemical (ICS) test is one among promising test, that we could use for detection of cervical precancerous changes and cancer in the future. For safe use Institute of Oncology Ljubljana developed a training program for the evaluation of p16/Ki-67 dual staining (DS) on conventional PAP smears. We aimed to test if this program is also suitable for teaching laboratory staff without skills in cervical cytology and immunocytochemistry. The training program was tested on two students without previous experience in gynecological cytology and immunocytochemistry. The training consisted of two parts. In first part students had short lecture about normal gynecological cytology and DS. After that they evaluated 118 slides with DS without possibility to consult SC. In the second part of training students first discussed cases with discordant results from first evaluation on multiheaded microspore with SC and afterwards they were tested with another 383 DS slides. However, in this part they could consult SC. All slides were reevaluated by SC. Exact agreement, Kappa statistics, sensitivity and specificity for CIN2+ were calculated for the students and SC and compared to reference (R). We also check how much time students use to evaluate one slide. The agreement between the evaluators and the R initial training was the best by SC in the additional training was better by Student 2 (S2). Sensitivity for CIN2+ decreased for both students with the increase of specificity after additional training. However, the sensitivity and specificity of R were still better than those of both students and also from SC. The difference was not significant. The results of S2 were the best among evaluators and were closest to the reference results. The best time for slide reading obtained S2 compared to S1 after additional training. After the training was completed S2 was competent for DS evaluation, while S1 would need additional education and had to work under supervision of SC.

• Keywords: p16, Ki-67, cervical cancer, immunocytochemistry, Human Papilloma Virus