HPV DNA chip The clinical significance of HPV DNA chip test in the management of Atypical Squamous Cells of Undetermined Significance 2003 1
HPV DNA chip The clinical significance of HPV DNA chip test in the management of Atypical Squamous Cells of Undetermined Significance 2003 1 2
. 2003 1 3
----------------------------------------------------- 5 Abstract ------------------------------------------------------- 7 List of tables ---------------------------------------------------- 9 I. ------------------------------------------------------- 10 II. ------------------------------------ 12 III. ----------------------------------------------- 15 IV. ----------------------------------------------------- 25 V. ------------------------------------------------------ 38 ---------------------------------------------------- 41 4
: ( ASCUS, Atypical squamous cells of undetermined significance ),. ASCUS, ASCUS HPV DNA chip. : 2001 11 1 2002 6 30 ASCUS, HPV DNA chip,, 48. HPV DNA chip 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68 69. : ASCUS 48 ( CIN II ) 9 18.8% 5
. 48 HPV DNA 20 41.7%. HPV DNA, 30.6%( 11/36 ), ( CIN I ) 33.3%( 1/3 ), ( CIN II ) 80%( 4/5 ), ( CIN III ) 100%( 2/2 ), 100%( 2/2 ). HPV DNA. HPV DNA chip, HPV DNA 88.9%, 69.2%, 96.4%, 40%. : ASCUS HPV DNA chip. 6
Abstract Objective: Cervical intraepithelial neoplasia ( CIN ) and invasive cervix cancer were detected in some case of Atypical squamous cells of undetermined significance ( ASCUS ) PAP smear. In this study, we attempted to assess the clinical significance of a cytologic diagnosis of ASCUS and determine the usefulness of HPV DNA chip test ( new diagnostic method for HPV ) in management and evaluation of ASCUS patients. Methods : This study was performed from 1. November 2001 to 30. June 2002 and included 48 cases of ASCUS who were evaluated by HPV DNA chip test and whose pathology was evaluated by punch biopsy, cone biopsy or hysterectomy. The result of type 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68 and 69 in HPV DNA chip test were categorized as high risk. Results: The detection rate of pathology of above CIN II in ASCUS was 18.8%( 9/48 ). The detection rate of high risk HPV DNA in ASCUS was 41.7%( 20/48 ). The detection rate of high risk HPV DNA in WNL or Reactive was 30.6%( 11/36 ), 33.3%( 1/3 ) in CIN I, 80%( 4/5 ) in CIN II, 100%( 2/2 ) in CIN III, 100%( 2/2 ) in invasive cervix cancer. Higher the grade of pathology, more the detection rate of high risk HPV DNA chip test. The sensitivity for 7
the prediction of pathology of above CIN II by HPV DNA chip test was 88.9% and specificity, negative predictive value, positive predictive value were 69.2%, 96.4% and 40%. Conclusion: The use of HPV DNA chip test in patients with ASCUS may provide usefulness in detection of pathology of CIN II, III and invasive cervix cancer. 8
List of tables Table 1. Age distribution of the patients of ASCUS Table 2. Pathologic diagnosis of ASCUS Table 3. High risk HPV detection rate in each pathologic diagnosis Table 4. Type of HPV in normal and CIN I patients Table 5. Type of HPV in CIN II, CIN III and invasive cancer patients Table 6. Correlation of pathologic diagnosis and high risk HPV DNA detection 9
I. 1943 Papanicolaou, 1., 6-55% 2. 1988 ( NCI ) The Bethesda system ( TBS ) ( ASCUS, Atypical squamous cells of undetermined significance ), ( LSIL ), ( HSIL ) 3. 2001 3 TBS ( Atypical squamous cells ) ASC-UC ( Atypical squamous cells of undetermined significance ) ASC-H ( Atypcial squamous cells, cannot exclude HSIL ). (ASC ) ( reactive ) 10
( SIL ) 5-13%,. ( human papilloma virus ).,... Hybrid Capture II.,, HPV DNA chip HPV DNA ASCUS. 11
II. 1. 2001 11 1 2002 6 30 ( ASCUS ), HPV DNA chip ( punch biopsy ), ( cone biopsy ), 48., HPV DNA chip,,. HPV DNA chip 1. 12
2. cytobrush. HPV speculum cytobrush ( ) Neodin TM PCR kit. HPV DNA Neodin TM PCR kit 22 HPV type specific oligonucleotide probe gamma-globin probe, probe DNA ( target DNA ) PCR ( hybridization ). probe streptavidin-r-phycoerythrin dye HPV. HPV DNA chip 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68 69 6, 11, 34,40, 42, 43 44.,,. 13
3. 48.,,,,,. HPV DNA chip. 4. Fisher's exact test P value 0.05. 14
III. 1. 48 44.8. 31 40 15 ( Table 1 ). 2. ASCUS ASCUS 48 36 (75.0%), ( CIN I ) 3 ( 6.2% ), ( CIN II ) 5 ( 10.4% ), ( CIN III ) 2 ( 4.2% ), 2 ( 4.2% ) ( Table 2 ). 48 9 18.8%. 15
3. HPV DNA 48 HPV DNA 20 41.7%. HPV DNA, 11 ( 30.6% ), 1 ( 33.3% ), 4 ( 80% ), 2 ( 100% ), 2 ( 100% ) ( Table 3). HPV DNA. HPV DNA 100%. 4. HPV DNA HPV DNA HPV DNA. HPV DNA 16 18, ( Table 4 ). 16
5. HPV DNA HPV DNA 16 37.5%, 18 12.5% ( Table 5 ). 6. HPV DNA,,. HPV DNA 39 27 69.2%, HPV DNA 39 12 30.8%. HPV DNA 9 1 11.1%, HPV DNA 9 8 88.9% ( Table 6 ). ASCUS HPV DNA chip 88.9%, 69.2%, 96.4%, 17
40%, HPV DNA P value 0.0049. 18
Table 1. Age distribution of the patients of ASCUS Age of patients No. of patients(%) 21-30 8 ( 16.7 ) 31-40 11 ( 22.9 ) 41-50 15 ( 31.2 ) 51-60 7 ( 14.6 ) 61-70 6 ( 12.5 ) 71-80 1 ( 2.1 ) Total 48 ( 100 ) 19
Table 2. Pathologic diagnosis of ASCUS* Pathologic diagnosis No. of patients(%) Normal or reactive 36 (75 ) CIN I ** 3 (6.2 ) CIN II 5 (10.4 ) CIN III 2 (4.2 ) Invasive cancer 2 (4.2 ) Total 48(100) *ASCUS : Atypical squamous cells of undetermined significance **CIN : Cervical intraepithelial neoplasia 20
Table 3. High risk HPV* detection rate in each pathologic diagnosis Pathologic diagnosis No.of cases No.of patients of high risk (%) Normal or reactive 36 11 (30.6 ) CIN I** 3 1 (33.3) CIN II 5 4 (80 ) CIN III 2 2 (100) Invasive cancer 2 2 (100) Total 48 20 (41.7 ) *HPV : Human papillomavirus **CIN : Cervical intraepithelial neoplasia 21
Table 4. Type of HPV in Normal and CIN I patients Case No HPV** DNA type Pathologic diagnosis 1 6* Normal or Reactive 2 16 Normal or Reactive 3 18 CIN I*** 4 40* Normal or Reactive 5 51 Normal or Reactive 6 56 Normal or Reactive 7 58 Normal or Reactive 8 58 Normal or Reactive 9 68 Normal or Reactive 10 51,58 Normal or Reactive 11 45,66 Normal or Reactive 12 66,68 Normal or Reactive 13 35,42 Normal or Reactive 14 16,18,34 Normal or Reactive *low risk type HPV **HPV: Human papillomavirus ***CIN : Cervical intraepithelial neoplasia 22
Table 5. Type of HPV in CIN II, CIN III and invasive cancer patients Case No HPV *DNA type Pathologic diagnosis 1 16 CIN II** 2 16 CIN II 3 16 Invasive cancer 4 18 CIN III 5 45 CIN II 6 35 CIN II 7 33,35 Invasive cancer 8 33,51 CIN III *HPV: Human papillomavirus **CIN: Cervical intraepithelial neoplasia 23
Table 6. Correlation of pathologic diagnosis and high risk HPV DNA detection High risk HPV* DNA Pathologic diagnosis CIN I** CIN II Total (%) (%) (%) Negative 27 ( 69.2 ) 1 ( 11.1 ) 28 ( 58.3 ) Positive 12 ( 30.8 ) 8 ( 88.9 ) 20 ( 41.7 ) Total 39 (100) 9 (100) 48(100) *HPV: Human papillomavirus **CIN: Cervical intraepithelial neoplasia 24
IV. 12%, 1.. CIN I, CIN II, CIN III, CIS,,..,,, HPV DNA,,,., Papanicolaou I, II, III, IV, V. 1943 25
Papanicolaou Traut,,., 1. 6-55% 2.,,.,,, 1988 Bethesda ( NCI ) PAP class II The Bethesda system ( TBS ). TBS,,,. TBS ASCUS, 26
( LSIL ), ( HSIL ) 3. HPV, ASCUS. ASCUS,,, HPV, koilocytotic atypia 4. 1988 TBS ASCUS ( Reactive ), ( SIL ), Papanicolaou class II,,,,, HPV.. ASCUS, 27
2-3,,,. 1991 TBS ASCUS fabor reactive ( ), fabor SIL ( ), NOS ( not otherwise specified ) 3. 2001 3 TBS ASCUS,,,, ASCUS., ASCUS ( ASC, Atypical squamous cell ). ASC SIL. ASC ASC-US ASC-H ASC-US ( Atypical squamous cells of undetermined significance ) SIL SIL ASCUS, fabor reactive ASCUS, NOS ASCUS, fabor SIL, ASC 95%. ASC-H ( Atypical squamous cells, cannot exclude HSIL ) HSIL HSIL 28
HSIL CIN II, CIN III ASC-US CIN II, CIN III ASC-US. ASC-H ASC 5-10%. ASC ASC 5%, SIL ASC ASC:SIL 2:1 3:1 5. ASCUS 2-11% 6., ASCUS 1997 2.1%, 1997 5.7%, 1993 1.6% 7.8.9.. ASCUS 1.98%. ASCUS ASCUS 46.2%, 53.8% 10. ASCUS 10, ASCUS 29
5-13% 11. ASCUS,,,,,,, HPV DNA 12., ASCUS 13.14. 9,. 2002 4 ASCCP ASC-US ASC-H, ASC-US,, HPV DNA. HPV DNA 30
, SIL, 15.16. ASC-H. 1960, 1980 ( HPV, Human papillomavirus ), HPV,, 17.18. 90%. 99% 1 HPV. HPV, HPV, HPV. HPV 6, 11, 42, 43, 44 condyloma, HPV 16, 18 31
, HPV 31, 33, 35, 39, 45, 51, 52, 56, 58 19. HPV E5, E7 HPV 18. HPV DNA 16 18, 31, 45., HPV HPV. HPV viral particle, HPV typing 100 HPV 20. 20 20. HPV HPV DNA HPV DNA. HPV DNA Liquid hybridization ( Hybrid capture system ) HPV type specific probe Dot blot hybridization, Southern blot hybridization, Filter in situ hybridization, HPV DNA 32
type specific PCR, general primer PCR, general primer set HPV DNA Dot blot hybridization, Microtiter plate hybridization, Line probe assay. HPV DNA, HPV DNA. HPV DNA chip HPV 22 type ( High risk group; 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 69 Low risk group; 6, 11, 34, 40, 42, 43, 44 ), HPV. HPV DNA,,,,,, scanner HPV DNA,. 33
HPV 3 HSIL. HPV HSIL,,. HPV borderline smear 21. HPV DNA. ASCUS HPV DNA LSIL, HPV DNA HSIL 22. Apgar ASCUS HPV DNA, HPV DNA,, 34
23. Cox ASCUS HPV DNA CIN II CIN III 93%, 27% 22. Hatch ASCUS CIN II CIN III HPV DNA 60% 24. HPV DNA ASCUS 25. Gideon ASCUS CIN II CIN III 24.6%., ASCUS HPV DNA CIN II CIN III 85.7%, 97%. HPV DNA CIN II CIN III 87%, 95.7% 24.6% 26. ASCUS HPV DNA CIN II 35
88.9%, 69.2% 41.7%. HPV, HPV DNA chip. ASCUS HPV DNA chip 6. Lorincz ASCUS DNA HPV, HPV DNA SIL, HPV DNA 6 HPV DNA 27. ASCUS HPV 36
22.24.28.,, HPV, HPV. ASCUS HPV DNA, 28.29. 6 HPV 30, HSIL HPV DNA 31. HPV DNA. 32.33.34. HPV DNA, 35. 37
V. 2001 11 1 2002 6 30 ASCUS, HPV DNA chip,, HPV DNA. 1. ASCUS 48 9 18.8%. ASCUS 18.8%. 2. 48 HPV DNA 20 41.7%. HPV DNA, 38
11 ( 30.6% ), 1 ( 33.3% ), 4 ( 80% ), 2 ( 100% ), 2 ( 100% ). HPV DNA. 3. HPV CIN II 16, 18. ASCUS. 4.,,. HPV DNA 39 27 69.2%, HPV DNA 39 12 30.8%. HPV DNA 9 1 11.1%, HPV DNA 9 8 88.9%. ASCUS HPV DNA chip 88.9%, 39
69.2%, 96.4%, 40%, HPV DNA P value 0.0049. ASCUS HPV DNA chip,. 16, 18. HPV DNA chip. 40
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