What is the proper technique for performing a Pap smear in a non‑pregnant woman aged 21‑65 with a cervix?

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How to Perform a Pap Smear

The Pap smear should be obtained using an Ayre spatula or similar collection device to sample the cervical transformation zone, with the specimen immediately spread on a glass slide and fixed in 95% ethyl alcohol, or placed in liquid-based cytology medium.

Patient Preparation and Positioning

  • Position the patient in the dorsal lithotomy position with appropriate draping for comfort and privacy 1, 2
  • Perform a speculum examination to visualize the cervix adequately before specimen collection 2
  • Ensure the patient has not had intercourse, douched, or used vaginal medications for 24-48 hours prior to the test for optimal specimen quality 1

Specimen Collection Technique

Sampling the Cervix

  • Use an Ayre spatula to sample the ectocervix by rotating it 360 degrees around the external os to capture cells from the transformation zone 1, 3, 2
  • For endocervical sampling, use a cytobrush or endocervical brush inserted into the endocervical canal and rotated 180-360 degrees 2
  • The transformation zone (squamocolumnar junction) is the critical area to sample, as this is where most cervical cancers and precancerous lesions arise 1

Specimen Handling

  • Immediately spread the collected material evenly across a clean, grease-free glass slide in a single stroke to avoid clumping 1, 3
  • Fix the slide immediately by immersing it in 95% ethyl alcohol fixative to prevent air-drying artifact 1, 3, 2
  • For liquid-based cytology, rinse the collection device in the provided vial containing preservative solution 4
  • Label the slide or vial with patient identification before fixation 2

Quality Assurance

  • An adequate sample should contain sufficient squamous cells and endocervical/transformation zone component 2
  • Studies show that 6.42% of samples may be inadequate, requiring repeat collection 2
  • Ensure proper technique to minimize inadequate specimens that delay diagnosis 2

Clinical Context for Screening

  • This technique applies to women aged 21-65 years undergoing routine cervical cancer screening 5, 4
  • For women 21-29 years, cytology alone every 3 years is recommended 5, 6, 4
  • For women 30-65 years, options include cytology every 3 years, primary HPV testing every 5 years, or co-testing every 5 years 7, 6, 4

Common Pitfalls to Avoid

  • Avoid air-drying the specimen before fixation, as this creates artifact that makes interpretation difficult 1, 3
  • Do not perform Pap smears during menses, as blood can obscure cellular detail 1
  • Ensure adequate sampling of the transformation zone, as sampling only the ectocervix may miss lesions 1, 2
  • Do not screen women under 21 years of age, as this leads to more harms than benefits 5, 4

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Guideline

Cervical Cancer Screening Guidelines

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2026

Guideline

Cervical Cancer Screening Guidelines

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

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Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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