@2025 Afarand., IRAN

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ISSN: 2251-8215    Sarem Journal of Reproductive Medicine 2017;1(4):165-169

Background
Cervical cancer is the second most common cancer in women, with an annual incidence of around half ‎a million people worldwide, leading to death in at least 50% of cases [1-3]. ... [4]. Human ‎papillomavirus (HPV) is known to be one of the most important causes of cervical cancer [5].‎
Previous Studies
‎... [5-7]. Human papillomavirus is a small, uncoated virus with a double-stranded DNA that has at least ‎‎100 different types [8]. Approximately 35 to 40 types of this virus infect epithelium of the skin or ‎mucous membrane in the female reproductive system [9, 10]. Most viral infections produce a specific, ‎undigested symptom and are spontaneously improved [8]. If the infection continues, because of the ‎high incidence of lesions in the virus and the self-limiting blemishes of the skin that often appear on ‎the palms and legs, malignant and invasive lesions appear to be a cervical cancer.‎‏ ‏Condyloma ‎acuminatum (anogenital warts) is one of the most commonly‏ ‏reported viral diseases transmitted ‎through sexual contact [8-11].‎ Mucosal types can be classified as high-risk and low-risk depending on the risk of the disease. Harmful ‎types of human papillomavirus play a major role in the development of cervical cancer [9]. Of these ‎viruses, at least 15 types of high-risk viruses cause more than 95% cervical cancer and 80% vaginal ‎cancer [1, 12]. Therefore, accurate diagnosis will have a significant impact on the control and ‎management of this cancer and promote the health of women in the community [13].‎ Currently, 118 HPV genotypes are categorized according to the type of biological wall and oncogenicity ‎and type of racial evolution. Types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73 and 82 are ‎known as high risk [14]. Types 26, 53, and 66 may have oncogenic properties [15]. An abnormal ‎epithelial cells of the cervix can be detected by staining a pap smear and observing it under a ‎microscope, but this diagnosis only applies to women with advanced disease risk, and about 50% of ‎lesions are not detected by Pap smear alone. Since the HPV virus does not grow in the culture medium ‎and serologic testing has limited accuracy (HPV infection produces a humoral immune response ‎against a capsid protein that remains antibodies for many years), serological tests are not appropriate ‎for the diagnosis of present and past infections [16]. As a result, the exact diagnosis of HPV infection ‎depends mainly on the molecular method. Molecular techniques can be used to detect and accurately ‎identify HPV, as well as to inform the patient and follow up after treatment [17].‎
Aim(s)
This study was conducted to determine genotypes of human papillomavirus in cervical fluid cytology.‎
Research type
This research is descriptive-cross sectional.
Research society, place and time
This study was carried out on cervical cytology samples from patients aged 25-50 years who referred ‎to Sarem hospital during one year from 2013 to 2014 who were diagnosed with papillomavirus and ‎confirmed by pathological and molecular studies. ‎
Sampling method and number
‎42 cytological samples of cervical fluid were studied. Criteria for entering the research included ‎women aged 25 to 50, satisfaction to participate in research, recruitment and confirmation of ‎papillomavirus infection by clinical and pathological examinations based on ICD-9.‎
Used devices&materials
Extracting DNA from samples was done using the High Pure PCR Template Preparation Kit (Roche).‎ Genotyping was performed on all HPV positive specimens using the HPV type 3.5 LCD Array (Chipron ‎GmbH, Berlin). In this method, a mixture of pre-labeled primers provided by the kit produces labeled ‎DNA viral compositions. The non-florescent labeled PCR parts are bonded to the dedicated probes ‎attached to the chips bottom with a hybridization buffer, and after washing the chips, the labeled ‎solution is poured onto them, and then the PCR components attached to the labels are seen by the ‎enzyme-substrate reaction in the form of blue spots. Data readout is done both by the scanner and by ‎comparison with the proposed model contained in the kit.‎ In this method, two pairs of primers were provided for PCR, a pair of primers MY09 / MY11 that are ‎commonly used, and the second pair, which produces a piece of 125 buffer pairs. Based on the ‎instructions of the kit, 5 μl of the two independent PCR products were mixed and used to continue ‎hybridization. By using the method used, 32 different types of HPV were identified that included 6, 11, ‎‎16, 18, 31, 35, 33, 39, 42, 44, 45, 51, 52, 56, 58, 59, 61, 62, 66, 67, 68, 70, 72, 73, 81, 82, 84, 90 and 91.‎
Findings by Text
Of the 42 LBC samples, 27 cases (64%) belonged to the high-risk group of the papillomavirus. Of these ‎‎27 samples, 14 cases (52%) had infections and 70.4% of the samples showed infections of type 16 and ‎‎18.‎ The most common type of HPV observed in samples was typed 16 with an abundance of 52%, ‎followed by typing 18 with 18.5%, typing 56, 58 and 66 each with a frequency of 11% and types 31 ‎and 33 with a frequency of 3.7%. Infection with different types of types was observed in 14 cases ‎‎(52%). In most cases, there were samples of type 16 infections.‎
Main comparison to the similar studies
‎... [18, 19]. Currently, vaccines against two types of HPV16 and HPV18 have been approved and used in ‎many countries [20, 21].‎ International epidemiological studies have shown that HPV16 and HPV18 are high-risk HPV viruses ‎and are seen as isolated and multi-viral infections in at least 75% of cervical cancers [2]. The two ‎viruses also account for 80% of vaginal and vulvar cancers, 92% of anal cancers, 95% of oral cancers ‎and 89% of throat cancer [12]. Six other types of virus (including 31, 33, 35, 45, 52 and 58) cause 19% ‎of cervical cancer, of which two types 45 and 31 are more likely to be detected than the rest and are ‎seen in 10% of cancers [2].‎ John hu et al., in their study of the effect of papilloma virus genotype on the severity and prognosis of ‎cervical neoplasia in patients, have shown that the type of virus also affects the severity of the disease. ‎In this study, it was stated that the spread of the cancerous area in the HPV types was significantly ‎different and the highest was related to type 18. Therefore, patients with cervical neoplasia with type ‎‎18 should be more carefully monitored than patients with other genotypes [22]. In the current study, ‎Type 18 was the second most common type of virus in the study population with a frequency of ‎‎18.5%.‎ In Iran, a study by Mahmoudi et al. to determine genotypes of human papillomavirus virus on cervical ‎cancer samples in Yazd province, the HPV genome was identified in 70% of the samples that types 16 ‎and 18 with the frequency of 70% and 16.7% respectively, were the most abundant genotype and other ‎identified types were 33, 45 and 73 [23]. In the study of the prevalence of human papillomavirus in 18-‎‎60 year-old married women with natural Pap smear referring to gynecologic clinics of Isfahan ‎University of Medical Sciences, in 46 HPV positive cases, there were type 16 in 15.18% (7 samples), ‎type 18 in 13.4% (6 samples) and type 11 or 6 in 21.74% (6 samples) [24]. Compared to the recent ‎research, Type 16 has been much less prevalent. In a comprehensive study of genotyping of 20,000 Pap smear genotypes in Kerman, 93.75% of HPV-‎positive people were diagnosed with type 16 and type18 was found only in 6.25% of them [25]. In ‎another study, in which Khoda karami et al. reviewed the prevalence of this infection in the areas of ‎Tehran, the prevalence of HPV was 7.8%, of which 5.1% was high risk [26].‎
Suggestions
According to various studies in this domain, it is suggested that by implementing a comprehensive ‎epidemiological plan in the country, the actual outbreak of the papillomavirus infection be determined, ‎and this be planned for prevention and control of this disease. It is also suggested that in further ‎research, the demographic, epidemiological and risk factors of the disease be investigated.‎
Limitations

Conclusions
Various types of HPV have been identified and the most common type is HPV16. Simultaneous ‎infections with several types of human papillomavirus are common in cervical cytological fluid ‎samples of positive people.‎
Acknowledgments

Conflict of interest

Ethical Permissions

Funding Sources