Combination analysis of a whole lymph node by one-step nucleic acid amplification and histology for intraoperative detection of micrometastasis.
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.
Taniyama
Kiyomi
K
Institute for Clinical Research, Department of Pathology, National Hospital Organization Kure Medical Center and Chugoku Cancer Center, Kure, Japan. taniyamak@kure-nh.go.jp
Motoshita
Junichi
J
Sakane
Junichi
J
Makita
Kyoko
K
Akai
Yasumasa
Y
Daito
Motonari
M
Otomo
Yasuhiro
Y
Ono
Hiroshi
H
Mizunoe
Tomoya
T
Takeuchi
Yukiyasu
Y
Tominaga
Harumi
H
Koseki
Masato
M
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
D014408:Biomarkers, Tumor; D053539:Keratin-19; D012333:RNA, Messenger; D012334:RNA, Neoplasm