title: Combination analysis of a whole lymph node by one-step nucleic acid amplification and histology for intraoperative detection of micrometastasis.
abstract: OBJECTIVE
  The aim was to develop a more efficient molecular detection system than histological examination (HE) for lymph node (LN) metastasis.
  
  
  METHODS
  Cytokeratin (CK) 19 mRNA copy numbers of 5 colon carcinoma cell lines (Lovo, DLD1, WiDr, Colo201 and Colo320) were calculated and compared by one-step nucleic acid amplification (OSNA) and conventional real-time reverse-transcription polymerase chain reaction (RT-PCR). Then, 91 LN submitted for HE from 6 patients with advanced colorectal adenocarcinoma and 64 LN submitted for frozen diagnosis from 47 patients with different malignancies were examined by OSNA and HE.
  
  
  RESULTS
  CK19 mRNA copy numbers of all but Colo320 cells detected by OSNA were within double of those detected by RT-PCR. The least cell count of Lovo cells detected at one reaction (2 microl) by OSNA was calculated as 0.8 cells. Carcinoma metastasis showing either HE+ or OSNA+ was detected in 7.9% of the LN from advanced colorectal adenocarcinomas and in 30.0% of the LN for frozen diagnosis from different malignancies; HE-/OSNA+ metastasis was detected in 4.8 and 4.0%, respectively. OSNA analysis of 1 LN could be completed within 40 min.
  
  
  CONCLUSIONS
  A combined analysis of LN by HE and OSNA could increase the sensitivity for detecting micrometastasis during surgery.
authors.affiliation: Institute for Clinical Research, Department of Pathology, National Hospital Organization Kure Medical Center and Chugoku Cancer Center, Kure, Japan. taniyamak@kure-nh.go.jp
authors.forename: Kiyomi
authors.initials: K
authors.lastname: Taniyama
authors.forename: Junichi
authors.initials: J
authors.lastname: Motoshita
authors.forename: Junichi
authors.initials: J
authors.lastname: Sakane
authors.forename: Kyoko
authors.initials: K
authors.lastname: Makita
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Akai
authors.forename: Motonari
authors.initials: M
authors.lastname: Daito
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Otomo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ono
authors.forename: Tomoya
authors.initials: T
authors.lastname: Mizunoe
authors.forename: Yukiyasu
authors.initials: Y
authors.lastname: Takeuchi
authors.forename: Harumi
authors.initials: H
authors.lastname: Tominaga
authors.forename: Masato
authors.initials: M
authors.lastname: Koseki
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis* / Q000378:metabolism / Q000556:secondary; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000378:metabolism; D003106:Colon / Q000473:pathology; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D005260:Female; D005629:Frozen Sections; D006801:Humans; D007432:Intraoperative Period; D053539:Keratin-19 / Q000235:genetics / Q000378:metabolism; D008197:Lymph Node Excision / Q000379:methods*; D008198:Lymph Nodes / Q000378:metabolism / Q000473:pathology*; D008207:Lymphatic Metastasis / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques; D012333:RNA, Messenger / Q000378:metabolism; D012334:RNA, Neoplasm / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D014408:Biomarkers, Tumor; D053539:Keratin-19; D012333:RNA, Messenger; D012334:RNA, Neoplasm