Pap and HPVDNA Before and After Acetic Acid
Condition: CIN
Study type: Observational

Summary: 
      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.
    

Detailed description: 
      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.
    

Observational model: Other
Time perspective: Cross-Sectional
Sampling method: Probability Sample
Study population: 
        Healthy Married women attending the Preventive Oncology screening clinic in the age group
        30 to 60 years for cervical cancer screening.
      
Biospecimen description: 
      HPV DNA TEST will be collected in stored as per protocol of storage and will be analysed by
      hybridisation method in microbiologt Dept of Tata Memorial hospital.
    
Keywords: Pap,HPVDNA