Pap and HPVDNA Before and After Acetic Acid
Pap and HPVDNA Before and After Acetic Acid
This is a pilot study to assess the cell adequacy and quality of Pap smears and adequacy of
 samples for HPV DNA virus test done after application of 5%acetic acid in a VIA test.Normally
 a Pap test and HPV DNA test is performed before application of acetic acid. It is assumed
 that application of acetic acid may alter the cell adequacy of Pap test and HPV DNA test, if
 any of the above tests is done immediately after VIA test.The present study aims to see the
 effect of 4-5% acetic acid [vinegar] on the cell adequacy and quality of Pap smear and sample
 adequacy for HPV DNA test.
Aim (of pilot study):

 To evaluate the cell adequacy and smear quality of Pap smears and adequacy of samples for HPV
 HC 2 collected before and after application of acetic acid.

 Objective of Pilot study-

 1. Agreement between the results of HC2 on cervical samples obtained from women before and
 5 to 10 mins after VIA test

 2. Correlation between RLU/ cutoff ratio of HC2 on cervical samples obtained from women
 before and 5 to 10 mins after VIA test.

 3. Agreement between the results of cytology on cervical smears obtained immediately before
 and 5 to 10 mins after VIA test from the same women.

 Methodology:

 Design: Pilot study

 Study Duration: Three months

 Study Site: Preventive Oncology screening clinic, Tata Memorial Hospital

 Sample Size calculation and power estimates:

 For the pilot study, sampling method is a convenience sample of 50 consecutive eligible women
 attending the Preventive Oncology screening clinic and consenting to participate in the
 study. These women will be randomly allocated using lottery method (wherein 25 chits with HPV
 and 25 chits with cytology written inside will be placed). For every participant woman a chit
 will be picked up and the woman will be allotted to either HPV or cytology testing depending
 on the message in the chit. Thereafter the chit will be discarded.

 In 25 samples Pap smears will be collected before and after application of acetic acid. Steps
 mentioned below.

 In rest 25 samples HPV DNA will be collected before and after application of acetic acid.
 Steps mentioned below.

 Step 1: A per speculum examination will be performed on eligible women attending the
 Preventive Oncology screening clinic and the findings of the same will be recorded.

 Step 2: Pap smear will be collected - Smear 1. OR Cervical cell samples will be collected for
 HPV HC II - HPV sample 1. Step 3: VIA will be performed and the findings will be noted. Step
 4: A second Pap smear will be collected 5 to 10 minutes after performing VIA when the woman
 is still on the examination table by the same provider- Smear 2.

 OR Cervical cell samples will be collected for HPV HC II - HPV sample 2. Step 5: The smears
 will be submitted to the cytology department for evaluation. The HPV HC II samples will be
 submitted to the Microbiology department for evaluation.

 Step 6: The cytologist reading the smear will be blinded as to which smear was taken before
 and which was taken after the application of acetic acid. The same cytologist will read the
 pre-VIA and the post-VIA smears.

 Similarly, the Microbiology department staff will be blinded as to which HPV sample was taken
 before and which was taken after the application of acetic acid. The paired samples will be
 analyzed at the same laboratory in the same batch of testing.

 Step 7: The screen negative women will undergo routine follow-up. The screen positive women
 will undergo Colposcopy, guided biopsy and further management according to the Tata Memorial
 Hospital protocol.

 Analysis of the variables Data entry will be done in the Department of Preventive Oncology,
 Tata Memorial Hospital using SPSS version 18. Checks for consistency, data safety and
 analysis will be carried out at regular intervals. Both descriptive and inferential
 statistics will be generated for describing variables under the study objectives. Agreement
 in HC2 results between pre-VIA and post-VIA samples will be estimated using kappa statistics.
 Similarly, agreement in pap results between pre-VIA and post-VIA samples will be estimated
 using kappa statistics. Sensitivity and specificity of HC2 test and pap test in detecting
 CIN2+ lesions will be calculated using negative colposcopy or biopsy as the gold standard and
 will be compared between the pre and post VIA samples.
Inclusion Criteria:

 -  Married women attending the Preventive Oncology screening clinic.

 -  women in age group 30 to 60 years

 Exclusion Criteria

 -  Unmarried women

 -  married women in the age group less than 30 years and above 60 years

 -  pregnant women

 -  women with history of total hysterectomy

 -  women with cervical cancer