Anal Injury Screening for High Risk HPV
Anal Injury Screening for High Risk HPV
The diagnosis of anal cancer is superior with the Anal Test system compared to liquid
 cytology and anoscopy.
DESIGN METHODOLOGY: Prospective, cross-sectional, comparative, double-blind, randomized.

 STUDY POPULATION:

 Group 1, 250 HIV negative patients (150 men and 100 women) from the oncology service who
 agree to participate in the study and with a negative diagnosis for HIV made at the
 Infectious Diseases Service.

 Group 2, 250 HIV positive patients (150 men and 100 women) attended by the Infectious Disease
 Service diagnosed with HIV positive. The sample size was adjusted to the donation of 500 Anal
 Test devices and the subsequent molecular determinations for each sample.

 PROCEDURE:

 In this study, men and women between 30 and 65 years old with active sexual life and who are
 susceptible to the development of anal cancer can participate.

 1. Immunosuppressed patients, a) infected with the human immunodeficiency virus, HIV or b)
 by medication after receiving an organ transplant.

 2. Individuals who have or have had anal intercourse. Women with a history of cervical,
 vulva, or vaginal cancer.

 PLANNED ACTIVITIES:

 First: Candidate patients who meet the inclusion criteria will have an explanation of the
 importance of their participation in this protocol, all their doubts will be resolved and
 they will be asked to sign the letter of informed consent with witnesses.

 Second: After signing the letter of consent, the service provider will make a questionnaire
 to know the clinical characteristics and identification of the patient, they will be assigned
 a consecutive number.

 If assignation is a non-number, the self-test will be performed as the first in sequence,
 followed by liquid cytology and then anoscopy.

 If assignation is an even number, the liquid cytology will be done as the first in sequence,
 followed by self-test and the anoscopy.

 -  For the Self-Test: the patient will be given the Anal Test medical device with the
 following recommendations: a) washing of patient hands, b) removing the plastic
 packaging from the device, c) removing the cover, d) Holding it by the handle, the
 device will be inserted into the anus, e) patient will be left intro-anal for 30 seconds
 and then it will be rotated 180 degrees, f) it will slowly withdraw the device and g) it
 will be delivered to the assistant nurse.

 -  The nurse will put the device (with the sample) in a tube with 5 ml of PreservCyt
 solution and close it tightly.

 -  The tube with the device and the sample will be kept at room temperature until transport
 to the laboratory for processing and obtaining results.

 -  The patient will answer a satisfaction questionnaire.

 -  An appointment will be given three weeks later to grant the result.

 Third:

 Processing of samples and issuance of results will be done in an external laboratory outside
 the Hospital General de México. In the Molecular Biology Laboratory, the coded samples will
 be processed as follows:

 1. DNA will be extracted using the Abbott Real-Time Polymerase Chain Reaction (RT-PCR)
 technology procedures, with the m2000 system. Presence and identification will be
 determined for HPV-Ar 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68.

 2. Only the positive samples from the first step; mRNA will be extracted and the viral
 oncology proteins expression of the E6 and E7 will be quantified for the following
 specific HPV-Ar 16, 18, 31, 33, 45, 52 and 58 by Real-Time Polymerase Chain Reaction
 (RT-PCR). The results of the laboratory studies will be sent to the responsible
 investigator.

 3. Processing of samples in an external laboratory outside the Hospital General de México
 and issuance of results. In the Molecular Biology Laboratory, the coded samples will be
 processed as follows:

 1. DNA will be extracted using the Abbott Real-Time Polymerase Chain Reaction (RT-PCR)
 technology procedures, with the m2000 system. The presence and identification will
 be determined for HPV-Ar 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68.

 2. Only the positive samples from the first step; mRNA will be extracted and the
 expression of the viral oncology proteins E6 and E7 will be quantified for the
 following specific HPV-Ar 16, 18, 31, 33, 45, 52 and 58 by Real-Time Polymerase
 Chain Reaction (RT-PCR). The results of the laboratory studies will be sent to the
 responsible investigator.

 Fourth:

 If they are positive to HPV-Ar, investigator will follow-up patient to corroborate the
 elimination or persistence of the HPV infection (surveillance). Patients who are positive for
 E6 and E7 messenger ribonucleic acid (mRNA) will be cited for high-resolution anoscopy and
 biopsy, negative patients will be under surveillance. Patients who are normal or with Low
 grade Intraepithelial Lesion (lSIL) (NIA1) by cytology will be under surveillance. High grade
 Intraepithelial Lesion (HSIL) patients (NIA2 and NIA3) will be scheduled for high-resolution
 anoscopy and biopsy.

 STATISTICAL ANALYSIS Statistical results will be reported first in a descriptive way,
 reporting means and standard deviations of the main study variables, as well as percentages
 of the presence of high-risk human papillomavirus analyzed in this investigation. Bi-variate
 comparative tests such as t-Student or Chi-square tests will be applied to identify possible
 differences between the group of people with the presence/absence of high-risk viruses and
 demographics and clinical variables such as the presence of HIV. When there is no normality,
 the non-parametric Mann-Whitney U test will be applied. Linear correlations will be analyzed
 with the Spearman and Pearson correlation tests depending on the type of analyzed variables.
 Data concordance analysis such as the kappa test will be performed because due to presence of
 human papillovirus HPV-Ar sub-types as well as lesion type compared to the results of liquid
 anal cytology that in this study the investigators will consider our "gold standard", in
 addition to perform sensitivity, specificity, positive and negative predictive value tests.
 Finally, and although it is not part of our main objectives, the investigators will analyze
 the possible associate risk factors with the presence of high-risk human papillovirus (HPV)
 using logistic regression models.

 ETHICAL ASPECTS AND BIOSECURITY. ETHICS. What is indicated in relation to ethics on human
 studies will be strictly followed according to Regulations of the General Law of Health in
 Matters of Health Research of Mexico, and the Declaration of Helsinki, considering subsequent
 versions.

 INFORMED CONSENT LETTER. SAFETY The subject under study will have complete information on the
 procedures that will be carrying out. The studies will be done exclusively with the
 assistance of the authorized research team and in consulting rooms belonging to the Oncology
 Service. If the subject under study expresses his desire not to participate in the process,
 it will be suspended immediately.

 All the studies to be carried out on the patient are totally safe and innocuous.
Inclusion Criteria:

 -  Men and women between 30 and 65 years old.

 -  They agree to participate in the study and sign informed consent.

 -  They have not received previous treatments for HPV or had previous intraepithelial
 lesions or anal cancer.

 -  Immunosuppressed patients.

 -  Women with LIAG of the genital tract (cervix, vagina, or vulva).

 -  Patients who have had anal intercourse

 Exclusion Criteria:

 -  Patients sexually inactive

 -  Subjects who had anal sex in less than 24 hours prior to the study.

 -  Patients with local medication application (enemas, suppositories).