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Taking a PAP SMEAR
Cervical Cancer : Pap smear
 George N Papanicolaou introduced cervical cytology in
clinical practice in 1940
 In 1945, PAP smear was endorsed by American cancer
society as an effective method for prevention of cervical
cancer
 Many countries now have National cervical screening
programs
Indian scenario
 Commonest cancer in women in India
 Major cause of deaths in women due to cancer
 Usually diagnosed at advanced stage
 No National program
 Uniformly low incidence of cervical screening in India
(6% in rich & 4% in poor)
Histological Types 30
 Squamous Cell Carcinoma : 80-95%
 Adenocarcinoma : 5-20%
 Other : Clear cell, sarcomas
Transformation zone
 Cervix develops from 2 embryonic sites
* from Mullerian duct - lined by columnar epithelium
* from urogenital plate - lined by stratified
squamous epithelium
 Point at which columnar and squamous epithelium
meet is called as original squamo-columnar
junction
Transformation zone
 Under influence of estrogen, original SCJ moves
onto the portio.
 Exposure of delicate columnar cells to vaginal
environment leads to squamous metaplasia.
 Transformation zone -
- Area of squamous metaplasia
- Area between original and new SCJ
Transformation zone
Transformation Zone -TZ
 Exposure of TZ to carcinogens begins the process of
intraepithelial neoplasia
 While exact role of carcinogens in this process remains poorly
understood, it is clear that HPV and cigarette smoking can
cause dysplasia at the TZ
 95% of cervical cancers develop in TZ
 Important to take sample from TZ
Transformation Zone
 Transformation zone may not be viewed during
routine speculum examination
Why cervical screening is a
feasible and useful strategy?
 Relative accessibility of cervix to take the smear
 Long natural history of cervical carcinogenesis
 Relative conservative treatment for premalignant lesions
 Cost effectiveness3
PAP Smear
 PAP smear sampling of cervix involves scraping of
cervical surface and a portion of non visualised
cervical canal using various sampling devices
Significance of Pap smear
 Detect precancerous & invasive cancer cervix cases in
early stages
 Positive screeners can be selected for selective tests and
management
 With treatment, progression of disease is halted. Thus
morbidity associated with advanced cancer decreases
 Mortality reduces by 20-60 %.
 Helps us to study natural history of disease.
Cervical Cancer : Pap smear
 Early detection of pre-malignant lesions by Pap
smears prevent at least 70% of potential cervical
cancers.
Of the 30% who actually
develop cervical cancer:
 8% elude cytological detection
- imperfections in cytological technology
- biologic behavior of malignant lesions
 22% represent women who develop cervical cancer because of
failure to regularly seek Pap smears => women whose cancers
could have been prevented with early detection and treatment.
How to take a Pap Smear ?
 Proper technique is very important
 More problems are due to improper sampling than
screening
 Not to be collected during menses
 Avoid vaginal contraceptives, vaginal medications for at
least 48 hrs before taking smear
 Abstinence for 24 hrs
 Postpartum smear should be taken only after 6 - 8 weeks of
delivery
 Patient in dorsal position
 Good illumination is necessary
 Cusco’s speculum is inserted to visualise & fix
the cervix
 Inspection of cervix done & findings are noted
 Ayres spatula is inserted first. It is placed at
cervical os so that longer end goes into cervical
canal and smaller end rests on ectocervix
How to take a Pap Smear ?
How to take a Pap Smear ?
 Spatula is rotated through 360 degrees
maintaining contact with ectocervix
 Do not use too much force [bleeding /pain]
 Do not use too less force [inadequate sample]
 Sample is smeared evenly on the slide and fixed
immediately
 Both sides of spatula are to be smeared
How to take a Pap Smear ?
 Endocervical sample is collected using an
endocervical brush
 Insert the cytobrush into canal, so that last bristles
of brush are visible
 Rotate the brush through 180 degrees. [more
rotations increase the chance of bleeding]
 Sample is rolled on the slide and fixed.
Fixation of smear
 Fixation is done immediately with
fixative like 95% alcohol or cytofix
spray to avoid air drying
 Spray should be kept at 10 inches, to
avoid destruction of cells by
propellent in the spray
 Smear should monolayer for proper
penetration of cell surface by fixative
How to take a Pap Smear ?
 Slide should be labeled properly with patients name,
identification no. and details
 Detailed history and clinical examination findings are to be
mentioned
 Patient details and clinical findings are to be maintained in a
register
 Advice is given regarding further follow up and treatment
Systems for cervical cytology reporting
 George N Papanicolaou (1954)
5 classifications based on certainty of finding malignant cells
 Descriptive system – WHO - (1968)
based on morphologic criteria – included mild, moderate,
severe dysplasia and Ca In Situ
 Richart – CIN –based on histologic diagnosis
Systems for cervical cytology reporting
 Bethesda system – TBS (1988)
National cancer institute revised in 1991 and 2001
 Adequacy of smear must be determined before reporting
Smear is adequate when
- Patient identification
- adequate clinical history
Bethesda system
 Interpretable cellular cytology
 not obscured by inflammation, debris, blood, drying
 not scanty smear
 Adequate sampling from transformation zone
 presence of at least 2 clusters of well preserved
endocervical cells or metaplastic cells
Bethesda system
Results :
 Within normal limits ( WNL )
Benign cellular changes - this term was removed and
group was included in WNL in 2001
 Reactive or Reparative changes – seen with atrophy,
inflammation, surgery, radiation, IUCD, tampoons
 Infections – trichomoniasis, fungal, bacterial, HSV.
Bethesda system - results
 Epithelial cells abnormalities
 Squamous cells
• ASCUS
• ASCUS-H - suggestive of high grade lesion
• LSIL - changes associated with HPV, atypical
changes, mild dysplasia/ CIN1
• HSIL – moderate to severe dysplasia / CIN2, 3
and Ca In Situ
• HSIL – where invasion cannot be ruled out
• Squamous cell carcinoma
Bethesda system
Results :
 Glandular cells – AGUS (Endocervical, endometrial)
Adenocarcinoma
(endocervical, endometrial, extrauterine)
 Other malignant neoplasms
Normal cervix-cytology
 Squamous cells
 Exfoliated indivisual cells
 Navicular in shape with abundant cytoplasm and small,
dark, round /oval, pyknotic nuclei
 Glandular cells
 Many times seen in clumps - linear or honeycombed
pattern.
 Slightly larger and basal nuclei
Cervical cytology - Inflammation
 Interpretation difficult due to inflammatory
background
 Lot of neutrophils and blood can obscure
cellular details
Low grade lesions
High grade lesions
High grade squamous
lesion
High grade glandular
lesion
Abnormal Pap smear- HPV
 Peripherial condensation of cytoplasm -
wire looping effect
 Koilocyte
PAP Descriptive CIN Bethesda
Class-1 negative negative WNL
Class 2
Inflammatory,
squamous, koilocytic
atypia
Reactive, reparatative
changes, ASCUS,
LSIL(HPV)
Class 3
Mild dysplasia
Moderate dysplasia
Severe dysplasia
CIN1
CIN2
CIN3
LSIL(HPV)
HSIL
HSIL
Class 4 Ca In Situ CIN3 HSIL
Class 5 Invasive Invasive Invasive
 Single test will not detect cervical abnormality but with 3
negative tests there is less than 1% chance of cervical
abnormality
 Conventional cytology has specificity of 98% and
sensitivity of 51%.
PAP smear
PAP Smears - Limitations
 Low sensitivity 51%
 False negative rates are due to faulty sampling, improper
fixation or interpretation problems
 Large group population & high risk group screening not
possible
 No consensus regarding testing
Pap smear as screening method
 New guidelines
 Target group - All women aged 18-70 yrs who have ever had sex
 Timing of Initial Screening -
Initial screening at age of 21 years or within 3 years of sexual activity
ACOG Guidelines-(Aug2003), American Cancer Society (Nov 2002) and
U.S. Preventative Services Task Force (Jan 2003)
Pap smear - guidelines
Screening interval - yearly till the age of 30 then 3 yearly
 When to End Screening
- After 70 yrs
- Post Hysterectomy
- done for benign lesions
- previous 3 normal PAP reports
- confirmed complete removal of cervical epithelium
Pap smear - guidelines
 In high risk group after treatment for CIN
every 3 monthly for 2 years
 every 6 monthly for 3yrs
 Yearly thereafter
 Women who had hysterectomy for CIN, it is necessary to do
vault smears
 In women who received vaccination against HPV, it is
necessary to continue screening
Liquid Based Cytology
 To improve results of PAP newer techniques like liquid
based cytology are recommended
 Cells are obtained with a broom, then the head is broken
off in to a vial containing preservative fluid
 In the laboratory the sample is spun to remove obscuring
material
 It gives clearer image, no cell clumps
 It will assist in future automated reading
 Several slides can be prepared from one smear
 Chlamydia, HPV testing can be done at later date
 Reduces the incidence of inadequate and repeat smears
Liquid Based Cytology
Cancer Cervix IS PREVENTABLE ,
IF Detected EARLY!!!!!!!!!
Thank You

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PAP test methods

  • 1. Taking a PAP SMEAR
  • 2. Cervical Cancer : Pap smear  George N Papanicolaou introduced cervical cytology in clinical practice in 1940  In 1945, PAP smear was endorsed by American cancer society as an effective method for prevention of cervical cancer  Many countries now have National cervical screening programs
  • 3. Indian scenario  Commonest cancer in women in India  Major cause of deaths in women due to cancer  Usually diagnosed at advanced stage  No National program  Uniformly low incidence of cervical screening in India (6% in rich & 4% in poor)
  • 4. Histological Types 30  Squamous Cell Carcinoma : 80-95%  Adenocarcinoma : 5-20%  Other : Clear cell, sarcomas
  • 5. Transformation zone  Cervix develops from 2 embryonic sites * from Mullerian duct - lined by columnar epithelium * from urogenital plate - lined by stratified squamous epithelium  Point at which columnar and squamous epithelium meet is called as original squamo-columnar junction
  • 6. Transformation zone  Under influence of estrogen, original SCJ moves onto the portio.  Exposure of delicate columnar cells to vaginal environment leads to squamous metaplasia.  Transformation zone - - Area of squamous metaplasia - Area between original and new SCJ
  • 8. Transformation Zone -TZ  Exposure of TZ to carcinogens begins the process of intraepithelial neoplasia  While exact role of carcinogens in this process remains poorly understood, it is clear that HPV and cigarette smoking can cause dysplasia at the TZ  95% of cervical cancers develop in TZ  Important to take sample from TZ
  • 9. Transformation Zone  Transformation zone may not be viewed during routine speculum examination
  • 10. Why cervical screening is a feasible and useful strategy?  Relative accessibility of cervix to take the smear  Long natural history of cervical carcinogenesis  Relative conservative treatment for premalignant lesions  Cost effectiveness3
  • 11. PAP Smear  PAP smear sampling of cervix involves scraping of cervical surface and a portion of non visualised cervical canal using various sampling devices
  • 12. Significance of Pap smear  Detect precancerous & invasive cancer cervix cases in early stages  Positive screeners can be selected for selective tests and management  With treatment, progression of disease is halted. Thus morbidity associated with advanced cancer decreases  Mortality reduces by 20-60 %.  Helps us to study natural history of disease.
  • 13. Cervical Cancer : Pap smear  Early detection of pre-malignant lesions by Pap smears prevent at least 70% of potential cervical cancers.
  • 14. Of the 30% who actually develop cervical cancer:  8% elude cytological detection - imperfections in cytological technology - biologic behavior of malignant lesions  22% represent women who develop cervical cancer because of failure to regularly seek Pap smears => women whose cancers could have been prevented with early detection and treatment.
  • 15. How to take a Pap Smear ?  Proper technique is very important  More problems are due to improper sampling than screening  Not to be collected during menses  Avoid vaginal contraceptives, vaginal medications for at least 48 hrs before taking smear  Abstinence for 24 hrs  Postpartum smear should be taken only after 6 - 8 weeks of delivery
  • 16.  Patient in dorsal position  Good illumination is necessary  Cusco’s speculum is inserted to visualise & fix the cervix  Inspection of cervix done & findings are noted  Ayres spatula is inserted first. It is placed at cervical os so that longer end goes into cervical canal and smaller end rests on ectocervix How to take a Pap Smear ?
  • 17. How to take a Pap Smear ?  Spatula is rotated through 360 degrees maintaining contact with ectocervix  Do not use too much force [bleeding /pain]  Do not use too less force [inadequate sample]  Sample is smeared evenly on the slide and fixed immediately  Both sides of spatula are to be smeared
  • 18. How to take a Pap Smear ?  Endocervical sample is collected using an endocervical brush  Insert the cytobrush into canal, so that last bristles of brush are visible  Rotate the brush through 180 degrees. [more rotations increase the chance of bleeding]  Sample is rolled on the slide and fixed.
  • 19. Fixation of smear  Fixation is done immediately with fixative like 95% alcohol or cytofix spray to avoid air drying  Spray should be kept at 10 inches, to avoid destruction of cells by propellent in the spray  Smear should monolayer for proper penetration of cell surface by fixative
  • 20. How to take a Pap Smear ?  Slide should be labeled properly with patients name, identification no. and details  Detailed history and clinical examination findings are to be mentioned  Patient details and clinical findings are to be maintained in a register  Advice is given regarding further follow up and treatment
  • 21. Systems for cervical cytology reporting  George N Papanicolaou (1954) 5 classifications based on certainty of finding malignant cells  Descriptive system – WHO - (1968) based on morphologic criteria – included mild, moderate, severe dysplasia and Ca In Situ  Richart – CIN –based on histologic diagnosis
  • 22. Systems for cervical cytology reporting  Bethesda system – TBS (1988) National cancer institute revised in 1991 and 2001  Adequacy of smear must be determined before reporting Smear is adequate when - Patient identification - adequate clinical history
  • 23. Bethesda system  Interpretable cellular cytology  not obscured by inflammation, debris, blood, drying  not scanty smear  Adequate sampling from transformation zone  presence of at least 2 clusters of well preserved endocervical cells or metaplastic cells
  • 24. Bethesda system Results :  Within normal limits ( WNL ) Benign cellular changes - this term was removed and group was included in WNL in 2001  Reactive or Reparative changes – seen with atrophy, inflammation, surgery, radiation, IUCD, tampoons  Infections – trichomoniasis, fungal, bacterial, HSV.
  • 25. Bethesda system - results  Epithelial cells abnormalities  Squamous cells • ASCUS • ASCUS-H - suggestive of high grade lesion • LSIL - changes associated with HPV, atypical changes, mild dysplasia/ CIN1 • HSIL – moderate to severe dysplasia / CIN2, 3 and Ca In Situ • HSIL – where invasion cannot be ruled out • Squamous cell carcinoma
  • 26. Bethesda system Results :  Glandular cells – AGUS (Endocervical, endometrial) Adenocarcinoma (endocervical, endometrial, extrauterine)  Other malignant neoplasms
  • 27. Normal cervix-cytology  Squamous cells  Exfoliated indivisual cells  Navicular in shape with abundant cytoplasm and small, dark, round /oval, pyknotic nuclei  Glandular cells  Many times seen in clumps - linear or honeycombed pattern.  Slightly larger and basal nuclei
  • 28. Cervical cytology - Inflammation  Interpretation difficult due to inflammatory background  Lot of neutrophils and blood can obscure cellular details
  • 30. High grade lesions High grade squamous lesion High grade glandular lesion
  • 31. Abnormal Pap smear- HPV  Peripherial condensation of cytoplasm - wire looping effect  Koilocyte
  • 32. PAP Descriptive CIN Bethesda Class-1 negative negative WNL Class 2 Inflammatory, squamous, koilocytic atypia Reactive, reparatative changes, ASCUS, LSIL(HPV) Class 3 Mild dysplasia Moderate dysplasia Severe dysplasia CIN1 CIN2 CIN3 LSIL(HPV) HSIL HSIL Class 4 Ca In Situ CIN3 HSIL Class 5 Invasive Invasive Invasive
  • 33.  Single test will not detect cervical abnormality but with 3 negative tests there is less than 1% chance of cervical abnormality  Conventional cytology has specificity of 98% and sensitivity of 51%. PAP smear
  • 34. PAP Smears - Limitations  Low sensitivity 51%  False negative rates are due to faulty sampling, improper fixation or interpretation problems  Large group population & high risk group screening not possible  No consensus regarding testing
  • 35. Pap smear as screening method  New guidelines  Target group - All women aged 18-70 yrs who have ever had sex  Timing of Initial Screening - Initial screening at age of 21 years or within 3 years of sexual activity ACOG Guidelines-(Aug2003), American Cancer Society (Nov 2002) and U.S. Preventative Services Task Force (Jan 2003)
  • 36. Pap smear - guidelines Screening interval - yearly till the age of 30 then 3 yearly  When to End Screening - After 70 yrs - Post Hysterectomy - done for benign lesions - previous 3 normal PAP reports - confirmed complete removal of cervical epithelium
  • 37. Pap smear - guidelines  In high risk group after treatment for CIN every 3 monthly for 2 years  every 6 monthly for 3yrs  Yearly thereafter  Women who had hysterectomy for CIN, it is necessary to do vault smears  In women who received vaccination against HPV, it is necessary to continue screening
  • 38. Liquid Based Cytology  To improve results of PAP newer techniques like liquid based cytology are recommended  Cells are obtained with a broom, then the head is broken off in to a vial containing preservative fluid  In the laboratory the sample is spun to remove obscuring material  It gives clearer image, no cell clumps  It will assist in future automated reading
  • 39.  Several slides can be prepared from one smear  Chlamydia, HPV testing can be done at later date  Reduces the incidence of inadequate and repeat smears Liquid Based Cytology
  • 40. Cancer Cervix IS PREVENTABLE , IF Detected EARLY!!!!!!!!! Thank You