PMID- 23968641
OWN - NLM
STAT- MEDLINE
DCOM- 20140108
LR  - 20221207
IS  - 1532-8392 (Electronic)
IS  - 0046-8177 (Linking)
VI  - 44
IP  - 11
DP  - 2013 Nov
TI  - Human papillomavirus multiplex ligation-dependent probe amplification assay for 
      the assessment of viral load, integration, and gain of telomerase-related genes 
      in cervical malignancies.
PG  - 2410-8
LID - S0046-8177(13)00228-1 [pii]
LID - 10.1016/j.humpath.2013.04.026 [doi]
AB  - We evaluated the reliability of a novel multiplex ligation-dependent probe 
      amplification (MLPA) assay in detecting integration of human papillomavirus (HPV) 
      based on the viral E2/E6 copy number ratio in formalin-fixed and 
      paraffin-embedded cervical lesions. The MLPA results were compared with those of 
      amplification of papillomavirus oncogene transcripts for RNA, detection of 
      integrated papillomavirus sequences for DNA, and HPV fluorescence in situ 
      hybridization (FISH). DNA was isolated from 41 formalin-fixed and 
      paraffin-embedded HPV-positive cervical lesions (cervical intraepithelial 
      neoplasia grade 3 lesions, squamous cell carcinomas, and adenocarcinomas) for 
      MLPA analysis. From 13 matching frozen samples, DNA and RNA were isolated for the 
      detection of integrated papillomavirus sequences and/or the amplification of 
      papillomavirus oncogene transcripts, respectively. Integrated HPV16, HPV18, or 
      both were identified. The MLPA assay detected viral integration in 12 of these 13 
      cases, and episomal copies also were detected in 7 cases. In 20 of the 24 cases 
      with exclusive viral integration or episomal viral copies as detected by FISH, 
      MLPA confirmed the physical status of the virus. In the cases classified as mixed 
      by FISH, the presence of excess episomal copies complicated the recognition of 
      viral integration by MLPA. Furthermore, the feasibility of detecting gain of the 
      telomerase genes with the HPV MLPA assay was evaluated. The MLPA confirmed the 
      FISH data in 12 of 13 cases in which the status of copy number gain for 
      telomerase RNA component was known. In conclusion, the HPV MLPA assay can be 
      performed on routinely processed cervical lesions for the detection of viral load 
      and HPV integration.
CI  - Copyright (c) 2013 Elsevier Inc. All rights reserved.
FAU - Theelen, Wendy
AU  - Theelen W
AD  - Department of Molecular Cell Biology, GROW - School for Oncology and 
      Developmental Biology, Maastricht University Medical Center, Maastricht 6200MD, 
      The Netherlands.
FAU - Litjens, Rogier J N T M
AU  - Litjens RJ
FAU - Vinokurova, Svetlana
AU  - Vinokurova S
FAU - Haesevoets, Annick
AU  - Haesevoets A
FAU - Reijans, Martin
AU  - Reijans M
FAU - Simons, Guus
AU  - Simons G
FAU - Smedts, Frank
AU  - Smedts F
FAU - Herrington, C Simon
AU  - Herrington CS
FAU - Ramaekers, Frans C S
AU  - Ramaekers FC
FAU - von Knebel Doeberitz, Magnus
AU  - von Knebel Doeberitz M
FAU - Speel, Ernst-Jan M
AU  - Speel EJ
FAU - Hopman, Anton H N
AU  - Hopman AH
LA  - eng
PT  - Comparative Study
PT  - Journal Article
DEP - 20130819
PL  - United States
TA  - Hum Pathol
JT  - Human pathology
JID - 9421547
RN  - 0 (DNA, Viral)
RN  - 0 (RNA, Viral)
RN  - 0 (telomerase RNA)
RN  - 63231-63-0 (RNA)
RN  - EC 2.7.7.49 (Telomerase)
SB  - IM
MH  - Carcinoma, Squamous Cell/pathology/*virology
MH  - Cervix Uteri/pathology/*virology
MH  - DNA, Viral/genetics
MH  - Female
MH  - Gene Dosage
MH  - Human papillomavirus 16/genetics/isolation & purification/physiology
MH  - Human papillomavirus 18/genetics/isolation & purification/physiology
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Multiplex Polymerase Chain Reaction/*methods
MH  - Papillomaviridae/genetics/*isolation & purification/physiology
MH  - Papillomavirus Infections/pathology/*virology
MH  - RNA/genetics
MH  - RNA, Viral/genetics
MH  - Reproducibility of Results
MH  - Telomerase/genetics
MH  - Uterine Cervical Neoplasms/pathology/*virology
MH  - Viral Load
MH  - Virus Integration
MH  - Uterine Cervical Dysplasia/pathology/*virology
OTO - NOTNLM
OT  - APOT
OT  - Cervical carcinoma
OT  - DIPS
OT  - HPV FISH
OT  - HPV integration
EDAT- 2013/08/24 06:00
MHDA- 2014/01/09 06:00
CRDT- 2013/08/24 06:00
PHST- 2012/12/23 00:00 [received]
PHST- 2013/03/26 00:00 [revised]
PHST- 2013/04/01 00:00 [accepted]
PHST- 2013/08/24 06:00 [entrez]
PHST- 2013/08/24 06:00 [pubmed]
PHST- 2014/01/09 06:00 [medline]
AID - S0046-8177(13)00228-1 [pii]
AID - 10.1016/j.humpath.2013.04.026 [doi]
PST - ppublish
SO  - Hum Pathol. 2013 Nov;44(11):2410-8. doi: 10.1016/j.humpath.2013.04.026. Epub 2013 
      Aug 19.