Rapid detection of metastasis of gastric cancer using reverse transcription loop-mediated isothermal amplification.
Tailor-made surgeries for patients with solid malignancies have been under consideration on the basis of the development of new approaches for minor metastatic foci of malignant tumors. Accurate and reliable methods to detect metastases in biopsy specimens with certain rapidity are essential for the performance of these surgeries. The aim of this study was to develop a rapid and practical method to detect metastasis in specimens from patients with gastric carcinoma with the use of reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction, a novel technique for detecting mRNA expressions of targeted sequences with high sensitivity, specificity and rapidity under isothermal conditions. RT-LAMP primers to detect cytokeratin19 (CK19) mRNA were generated and 92 lymph nodes (LNs) obtained from 9 patients with gastric cancer were tested for tumor metastases with this technique. Among 92 LNs, 15 were metastasis-positive by routine histopathological examination. RT-LAMP reaction detected CK19 expression in all of the pathologically positive LNs and in 16 of 77 negative LNs. Nested RT-PCR assay for CK19 expression was also performed on 2 of the 9 cases including 32 LNs. The agreement rate of CK19 expression detection by RT-LAMP and RT-PCR analysis was 31/32 (97%). The RT-LAMP technique showed similar sensitivity to detect metastases as nested RT-PCR assay, with a rapidity comparable to that of intraoperative histopathological examination with frozen sectioning and hematoxylin and eosin staining. This method is expected to play an essential role in the performance of tailor-made surgeries in the near future.
Horibe
Daisuke
D
Department of Frontier Surgery, Graduate School of Medicine, Chiba University, Chiba, Japan.
Ochiai
Takenori
T
Shimada
Hideaki
H
Tomonaga
Takeshi
T
Nomura
Fumio
F
Gun
Ma
M
Tanizawa
Tohru
T
Hayashi
Hideki
H
D000368:Aged; D001483:Base Sequence; D002277:Carcinoma / Q000175:diagnosis* / Q000556:secondary*; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D053539:Keratin-19 / Q000235:genetics*; D008198:Lymph Nodes / Q000737:chemistry / Q000473:pathology*; D008207:Lymphatic Metastasis; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013274:Stomach Neoplasms / Q000175:diagnosis / Q000473:pathology*
D053539:Keratin-19; D012333:RNA, Messenger