Background: Pitfalls in Pap test could be defined as false positive, false negative, or underdiagnosed results which can lead to unnecessary diagnostic procedures or delayed and inadequate treatment. It can be a consequence of misinterpretation of certain morphological entities which are described in this paper. Summary: The paper presents an overview of the morphological features and look-alikes of the common sources of pitfalls such as atrophy, repair, intrauterine device change, tubal metaplasia, hyperchromatic crowded groups, and radiation changes. Rare causes of pitfalls such as Arias-Stella changes, pemphigus, tumor diathesis per se, rare types of cervical cancer, including verrucous and papillary squamous cell cancer, gastric type, and endometrioid adenocarcinoma are also described. Key Messages: The awareness of pitfalls in cervical cytology is important for cytopathologists and clinicians to avoid future errors. Review of Pap tests with erroneous diagnosis is important for quality control in cytology laboratory, and it must be considered an educational- and experience-building procedure. Cytopathologist should not pull back in significant diagnoses, especially in human papillomavirus-negative cases.

1.
Bruni
L
,
Serrano
B
,
Roura
E
,
Alemany
L
,
Cowan
M
,
Herrero
R
, et al
.
Cervical cancer screening programmes and age-specific coverage estimates for 202 countries and territories worldwide: a review and synthetic analysis
.
Lancet Glob Health
.
2022
;
10
(
8
):
e1115
27
. Erratum in: Lancet Glob Health. 2023 Jul;11(7):e1011.
2.
Koliopoulos
G
,
Nyaga
VN
,
Santesso
N
,
Bryant
A
,
Martin-Hirsch
PP
,
Mustafa
RA
, et al
.
Cytology versus HPV testing for cervical cancer screening in the general population
.
Cochrane Database Syst Rev
.
2017
;
8
(
8
):
CD008587
.
3.
Terasawa
T
,
Hosono
S
,
Sasaki
S
,
Hoshi
K
,
Hamashima
Y
,
Katayama
T
, et al
.
Comparative accuracy of cervical cancer screening strategies in healthy asymptomatic women: a systematic review and network meta-analysis
.
Sci Rep
.
2022
;
12
(
1
):
94
.
4.
Nayar
R
,
Wilbur
DC
, editors.
The Bethesda system for reporting cervical cytology. Definitions, criteria, and explanatory notes
. 3rd ed.
New York
:
Springer
;
2015
.
5.
Torous
VF
,
Pitman
MB
.
Interpretation pitfalls and malignant mimics in cervical cytology
.
J Am Soc Cytopathol
.
2021
;
10
(
2
):
115
27
.
6.
Shin
E
,
Yu
J
,
Hong
SW
.
Trouble-makers in cytologic interpretation of the uterine cervix
.
J Pathol Transl Med
.
2023
;
57
(
3
):
139
46
.
7.
Sirota
RL
.
Defining error in anatomic pathology
.
Arch Pathol Lab Med
.
2006
;
130
(
5
):
604
6
.
8.
Kahn
JA
,
Slap
GB
,
Bernstein
DI
,
Tissot
AM
,
Kollar
LM
,
Hillard
PA
, et al
.
Personal meaning of human papillomavirus and Pap test results in adolescent and young adult women
.
Health Psychol
.
2007
;
26
(
2
):
192
200
.
9.
Jentschke
M
,
Lehmann
R
,
Drews
N
,
Hansel
A
,
Schmitz
M
,
Hillemanns
P
.
Psychological distress in cervical cancer screening: results from a German online survey
.
Arch Gynecol Obstet
.
2020
;
302
(
3
):
699
705
.
10.
Tambouret
RH
,
Wilbur
DC
.
The many faces of atrophy in gynecologic cytology
.
Clin Lab Med
.
2003
;
23
(
3
):
659
79
.
11.
Saminathan
T
,
Lahoti
C
,
Kannan
V
,
Kline
TS
.
Postmenopausal squamous-cell atypias: a diagnostic challenge
.
Diagn Cytopathol
.
1994
;
11
(
3
):
226
30
.
12.
Colgan
TJ
,
Woodhouse
SL
,
Styer
PE
,
Kennedy
M
,
Davey
DD
.
Reparative changes and the false-positive/false-negative Papanicolaou test: a study from the college of American pathologists interlaboratory comparison program in cervicovaginal cytology
.
Arch Pathol Lab Med
.
2001
;
125
(
1
):
134
40
.
13.
Crothers
BA
,
Booth
CN
,
Darragh
TM
,
Zhao
C
,
Souers
RJ
,
Thomas
N
, et al
.
False-positive Papanicolaou (PAP) test rates in the College of American Pathologists PAP education and PAP proficiency test programs: evaluation of false-positive responses of high-grade squamous intraepithelial lesion or cancer to a negative reference diagnosis
.
Arch Pathol Lab Med
.
2014
;
138
(
5
):
613
9
.
14.
Staats
PN
,
Souers
RJ
,
Nunez
AL
,
Li
Z
,
Kurtycz
DFI
,
Goodrich
K
, et al
.
The differential diagnosis of reparative changes and malignancy: performance in the college of American pathologists Pap education and proficiency testing programs
.
Arch Pathol Lab Med
.
2020
;
144
(
7
):
846
52
.
15.
Zhao
C
,
Crothers
BA
,
Tabatabai
ZL
,
Li
Z
,
Ghofrani
M
,
Souers
RJ
, et al
.
False-negative interpretation of adenocarcinoma in situ in the college of American pathologists gynecologic PAP education program the college of American pathologists gynecologic PAP education program
.
Arch Pathol Lab Med
.
2017
;
141
(
5
):
666
70
.
16.
Kamal
MM
.
The Pap smear in inflammation and repair
.
CytoJournal
.
2022
;
19
:
29
.
17.
Selvaggi
SM
,
Haefner
HK
.
Microglandular endocervical hyperplasia and tubal metaplasia: pitfalls in the diagnosis of adenocarcinoma on cervical smears
.
Diagn Cytopathol
.
1997
;
16
(
2
):
168
73
.
18.
Rintala
MA
,
Rantanen
VT
,
Salmi
TA
,
Klemi
PJ
,
Grénman
SE
.
PAP smear after radiation therapy for cervical carcinoma
.
Anticancer Res
.
1997
;
17
(
5B
):
3747
50
.
19.
Poflee
SV
,
Bhatia
JK
.
Cervical cytology: radiation and other therapy effects
.
CytoJournal
.
2022
;
19
:
32
.
20.
Benoit
JL
,
Kini
SR
.
“Arias-Stella reaction”-like changes in endocervical glandular epithelium in cervical smears during pregnancy and postpartum states-a potential diagnostic pitfall
.
Diagn Cytopathol
.
1996
;
14
(
4
):
349
55
.
21.
Lui
M
,
Boerner
S
.
Arias-Stella reaction in a cervicovaginal smear of a woman undergoing infertility treatment: a case report
.
Diagn Cytopathol
.
2005
;
32
(
2
):
94
6
.
22.
Meisels
A
,
Ayotte
D
.
Cells from the seminal vesicles: contaminants of the V-C-E smear
.
Acta Cytol
.
1976
;
20
(
3
):
211
9
.
23.
Wojcik
EM
,
Bassler
TJ
Jr
,
Orozco
R
.
DNA ploidy in seminal vesicle cells. A potential diagnostic pitfall in urine cytology
.
Anal Quant Cytol Histol
.
1999
;
21
(
1
):
29
34
.
24.
Lobo
AZ
,
Sousa
JX
Jr
,
Aoki
V
,
Sotto
MN
,
Aida Gay de Pereyra
E
,
Maruta
CW
, et al
.
Pemphigus vulgaris of the uterine cervix: misinterpretation of Papanicolaou smears
.
Acta Derm Venereol
.
2006
;
86
(
4
):
355
6
.
25.
Chivukula
M
,
Austin
RM
,
Shidham
VB
.
Evaluation and significance of Hyperchromatic Crowded Groups (HCG) in liquid-based paps
.
CytoJournal
.
2007
;
4
:
2
.
26.
Croll
E
,
Rana
DN
,
Walton
LJ
.
Hyperchromatic crowded cell groups in gynaecological liquid-based cytology samples
.
Br J Biomed Sci
.
2010
;
67
(
3
):
154
65
. quiz 165.
27.
Torous
VF
.
Challenging lesions in cervical cytology: the elusive HSIL
.
Cytopathology
.
2024
;
35
(
1
):
48
59
.
28.
Šekoranja
D
,
Repše Fokter
A
.
Triaging atypical squamous cells-cannot exclude high-grade squamous intraepithelial lesion with p16/ki67 dual stain
.
J Low Genit Tract Dis
.
2017
;
21
(
2
):
108
11
.
29.
Gajsek
US
,
Dovnik
A
,
Takac
I
,
Ivanus
U
,
Jerman
T
,
Zatler
SS
, et al
.
Diagnostic performance of p16/Ki-67 dual immunostaining at different number of positive cells in cervical smears in women referred for colposcopy cervical smears in women referred for colposcopy
.
Radiol Oncol
.
2021
;
55
(
4
):
426
32
.
30.
Peeters
E
,
Wentzensen
N
,
Bergeron
C
,
Arbyn
M
.
Meta-analysis of the accuracy of p16 or p16/Ki-67 immunocytochemistry versus HPV testing for the detection of CIN2+/CIN3+ in triage of women with minor abnormal cytology triage of women with minor abnormal cytology
.
Cancer Cytopathol
.
2019
;
127
(
3
):
169
80
,
31.
Negri
G
,
Bellisano
G
,
Carico
E
,
Faa
G
,
Kasal
A
,
Antoniazzi
S
, et al
.
Usefulness of p16ink4a, ProEX C, and Ki-67 for the diagnosis of glandular dysplasia and adenocarcinoma of the cervix uteri glandular dysplasia and adenocarcinoma of the cervix uteri
.
Int J Gynecol Pathol
.
2011
;
30
(
4
):
407
13
.
32.
Wright
TC
Jr
,
Stoler
MH
,
Ranger-Moore
J
,
Fang
Q
,
Volkir
P
,
Safaeian
M
, et al
.
Clinical validation of p16/Ki-67 dual-stained cytology triage of HPV-positive women: results from the IMPACT trial
.
Int J Cancer
.
2022
;
150
(
3
):
461
71
.
33.
Clark
SB
,
Dawson
AE
.
Invasive squamous-cell carcinoma in ThinPrep specimens: diagnostic clues in the cellular pattern
.
Diagn Cytopathol
.
2002
;
26
(
1
):
1
4
.
34.
Barrigón
A
,
Ziadi
S
,
Jacot-Guillarmod
M
,
Da Silva
S
,
Dumont
M
,
Raineri
I
, et al
.
Nabothian cyst content: a potential pitfall for the diagnosis of invasive cancer on Pap test cytology
.
Diagn Cytopathol
.
2019
;
47
(
2
):
127
9
.
35.
Pantanowitz
L
,
Upton
MP
,
Wang
HH
,
Nasser
I
.
Cytomorphology of verrucous carcinoma of the cervix. A case report
.
Acta Cytol
.
2003
;
47
(
6
):
1050
4
.
36.
Randall
ME
,
Andersen
WA
,
Mills
SE
,
Kim
JA
.
Papillary squamous cell carcinoma of the uterine cervix: a clinicopathologic study of nine cases
.
Int J Gynecol Pathol
.
1986
;
5
(
1
):
1
10
.
37.
WHO Classification of Tumours Editorial Board
.
World health organization classification of tumours
.In:
Female genital tumours
. 5th ed.
Lyon
:
IARC Press
;
2020
.
38.
Eleutério
JJ
,
Giraldo
PC
,
Cavalcante
DIM
,
Gonçalves
AKS
,
Eleutério
RMN
,
Giraldo
HPD
.
Papillary squamous cell carcinoma of the uterine cervix, high-risk human papilloma virus infection and p16(INK4a) expression: a case report Virus Infection and p16INK4a Expression. A Case Report
.
Acta Cytol
.
2009
;
53
(
2
):
188
90
.
39.
Turker
LB
,
Gressel
GM
,
Abadi
M
,
Frimer
M
.
Papillary squamous cell carcinoma of the cervix: two cases and a review of the literature
.
Gynecol Oncol Rep
.
2016
;
18
:
18
21
.
40.
Wilbur
DC
.
Practical issues related to uterine pathology: in situ and invasive cervical glandular lesions and their benign mimics: emphasis on cytology-histology correlation and interpretive pitfalls
.
Mod Pathol
.
2016
;
29
(
Suppl 1
):
S1
11
.
41.
Conrad
RD
,
Liu
AH
,
Wentzensen
N
,
Zhang
RR
,
Dunn
ST
,
Wang
SS
, et al
.
Cytologic patterns of cervical adenocarcinomas with emphasis on factors associated with underdiagnosis
.
Cancer Cytopathol
.
2018
;
126
(
11
):
950
8
.
42.
Lu
S
,
Shen
D
,
Zhao
Y
,
Kang
N
,
Wang
X
.
Primary endocervical gastric-type adenocarcinoma: a clinicopathologic and immunohistochemical analysis of 23 cases
.
Diagn Pathol
.
2019
;
14
(
1
):
72
.
43.
Schwock
J
,
Starova
B
,
Khan
ZF
,
Mirkovic
J
,
Parra-Herran
C
,
Ko
HM
, et al
.
Cytomorphologic features of gastric-type endocervical adenocarcinoma in liquid-based preparations
.
Acta Cytol
.
2021
;
65
(
1
):
56
66
.
44.
Ryu
A
,
Nagata
S
,
Kubo
C
,
Ueda
Y
,
Tanada
S
,
Idota
A
, et al
.
Conventional direct smear yields diagnostic indicators of gastric-type mucinous carcinoma compared with cytomorphological features identified by liquid-based cervical cytology
.
Acta Cytol
.
2021
;
65
(
2
):
150
7
.
45.
Serdy
K
,
Yildiz-Aktas
I
,
Li
Z
,
Zhao
C
.
The value of Papanicolaou tests in the diagnosis of endometrial carcinoma: a large study cohort from an academic medical center
.
Am J Clin Pathol
.
2016
;
145
(
3
):
350
4
.
46.
de Peralta-Venturino
MN
,
Purslow
MJ
,
Kini
SR
.
Endometrial cells of the “Lower Uterine Segment” (LUS) in cervical smears obtained by endocervical brushings: a source of potential diagnostic pitfall
.
Diagn Cytopathol
.
1995
;
12
(
3
):
263
71
; discussion 268-71.
47.
Lundeen
SJ
,
Horwitz
CA
,
Larson
CJ
,
Stanley
MW
.
Abnormal cervicovaginal smears due to endometriosis: a continuing problem
.
Diagn Cytopathol
.
2002
;
26
(
1
):
35
40
.
48.
Arbyn
M
,
Anttila
A
,
Jordan
J
,
Ronco
G
,
Schenck
U
,
Segnan
N
, et al
.
European guidelines for quality assurance in cervical cancer screening. Second edition: summary document
.
Ann Oncol
.
2010
;21(
3
):
448
58
.
49.
Kamal
M
.
Pap smear collection and preparation: key points
.
CytoJournal
.
2022
;
19
:
24
.
50.
Eversole
GM
,
Moriarty
AT
,
Schwartz
MR
,
Clayton
AC
,
Souers
R
,
Fatheree
LA
, et al
.
Practices of participants in the college of american pathologists interlaboratory comparison program in cervicovaginal cytology, 2006
.
Arch Pathol Lab Med
.
2010
;
134
(
3
):
331
5
.
51.
Nguyen
LN
,
Crothers
BA
,
Davey
DD
,
Natale
KE
,
Nunez
AL
,
Harkcom
T
, et al
.
Current state of cytologic-histologic correlation implementation for north American and international laboratories: results of the college of American pathologists cytopathology committee laboratory practices in gynecologic cytology survey
.
Arch Pathol Lab Med
.
2023
;
147
(
1
):
52
61
.
52.
Horback
K
,
Sundling
KE
,
Schmidt
RJ
,
Cibas
ES
.
Developing dashboards for performance improvement in cytopathology
.
J Am Soc Cytopathol
.
2021
;
10
(
6
):
535
42
.
You do not currently have access to this content.