Chromogen labeled in situ hybridization (CISH) approach for detection of high risk types human papillomavirus in liquid based cytology preparation of cervical smear in lraqi women

Background, human papillomavirus (HPV) is the most important factor in cervical carcinogenesis, and infection by the virus seems to be an early event in malignant transformation. High-risk human papillomaviruses (HPV) play the leading etiological role in the development of cervical, anal, and vaginal cancers and a substantial proportion of penile, vulvar, and oropharyngeal (tonsillar) cancers. High-risk types (currently including types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 69, 73, 82) can cause low-grade cervical cell abnormalities, high-grade cervical cell abnormalities that are precursors to cancer, and anogenital cancers. High-risk HPV types are detected in 99% of cervical cancers. Transmission of HPV infection occurs through skin-to-skin contact, in the case of genital HPVs this transition takes place as a result of sexual activities. Cervical cancer continues to cause significant morbidity and mortality over the past few decades. After breast cancer, cervical cancer is the second most common type of cancer in women worldwide. Materials and methods, this study was designed as a two prospective groups. It involved (80) selected Pap smear samples taken from women cervix that divided into two groups: The first group: include 50 Pap smear of malignant cervical cases as study group. The second group: include 30 Pap smear of nonmalignant as control study. all specimens (pap smear slides) were collected from department cytology –General Health Laboratory that belong to Ministry of health Central Public Health Laboratories and Al-Sadr General Hospital in Maysan Governorate in Iraq from period (2019) to (2023) ,The diagnosis of these slides were made using standard cytological criteria ,these slides re-examined by chromogenic in situ hybridization technique (CISH) after destaining slides by rehydration (alcohols series). The practical work for this study conducted the followings: Remove cover slides by soaking in xylene. Destaining of slides by rehydration series (alcohol series). Fixing slides in 1% formaldehyde and then dry. Applying detection system and probe. Evaluation the H-R HPV signals by light microscope. Results, The age of study groups ranged from 20 to ≥50 years old, cervical carcinoma was increased in age interval (40 - 49) which was (28 %) followed by ( 24%) of the cases distributed in age interval (≥50) followed by (22%) CISH signals for HR-HPVs DNA were monitored in (90%) of cervical carcinoma cases. It was higher than that found in control group which made (16.7%). Significant difference was found between CISH results and cytology examinations were monitored in (57.8%) in ASCUS distributed among age groups, followed by (15.6%) in HSIL, followed by (11.1%) in LSIL, followed by (8.9%) in cervicitis, followed by (4.4%) in ASCH, followed by (2.2) in atrophy and finally (0%) for normal. Non-significant difference between CISH results and cytology examination. Significant difference between CISH results and number of abortion. Furthermore, patterns of replication showed that (26%) of cervical carcinoma cases were punctuated (integrated DNA), followed by (24%) (which involved integrated as well as episomal HR-HPVs DNA), followed by (20%) was diffused (episomal DNA) in case study, while only (17%) in control study was diffused. Non significant different between CISH results and number of delivery. Conclusion, Our results confirm that an HPV CISH assay is reasonably sensitive for HR-HPVs infection and that it might be useful to identify integrated HPV DNA in cell specimens, The standards for HPV detection in cytological specimens, CISH stain technologies would be considered a viable potential alternative to molecular assays in the evaluation of the biology of cervical lesions.