Rapid diagnosis of lymph node metastasis in lung cancer with loop-mediated isothermal amplification assay using carcinoembryonic antigen–mRNA

Maeda, Jun; Inoue, Masayoshi; Nakabayashi, Kadzuki; Otomo, Yasuhiro; Shintani, Yasushi; Ohta, Mitsunori; Okumura, Meinoshin; Matsuura, Nariaki

doi:10.1016/j.lungcan.2008.12.003

Cited by 12 publications

(8 citation statements)

References 23 publications

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“…As a consequence, seven out of 10 patients studied were upstaged according to TNM classification (data not shown). These results are comparable with reports on RT-LAMP analysis for nodal metastasis of other malignancies (25,26), showing relatively lower sensitivities compared with quantitative RT-PCR analysis with the same malignancies (10). Meanwhile, relatively higher false positivity was observed in this analysis (9% for nodes from patient with gastritis).…”

Section: Discussionsupporting

confidence: 86%

Rapid diagnosis of micrometastasis of gastric cancer using reverse transcription loop-mediated isothermal amplification

Muto

Matubara²,

Tanizawa³

et al. 2011

Oncol Rep

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Abstract. Methods to detect metastases in biopsy specimens with certain rapidity and accuracy are essential to performing tailor-made surgeries for solid malignancies. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction is a novel technique for detecting mRNA expression of target sequences with high sensitivity and rapidity, even from crude samples without RNA purification. Applicability to detect lymph node (LN) micrometastasis of gastric cancer was tested. Total of 26 LNs were retrieved from 10 patients with primary gastric cancer. Each LN was serially sectioned, and every set of three serial sections were tested for routine histopathological (H&E) and immunohistochemical examination with anti-cytokeratin antibodies (IHC), and RT-LAMP analysis targeted cytokeratin 19 mRNA. Results from H&E/IHC and RT-LAMP analysis were compared in each set of sections. All the sections of those containing metastatic lesions equivalent to a volume of overt metastasis (maximum diameter >2 mm), 90% of those containing micrometastasis (between 2 and 0.2 mm) and 83% of those containing isolated tumor cells (<0.2 mm) were detectable using this procedure. Total analysis from lysates of clinical specimens required <75 min. This new technique is suggested to be an alternative to rapid diagnosis of micrometastasis based on conventional histopathological analysis.

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Section: Discussionsupporting

confidence: 86%

Rapid diagnosis of micrometastasis of gastric cancer using reverse transcription loop-mediated isothermal amplification

Muto

Matubara²,

Tanizawa³

et al. 2011

Oncol Rep

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“…In this setting, the development of the loop-mediated isothermal amplification method (which was recently introduced as a fast diagnostic procedure for infectious disorders) may prove significant as it has been successfully tested for detecting CEA mRNA in intra-thoracic lymph nodes. Such new rapid procedures may be proposed during mediastinoscopy or transbronchial preoperative lymph node staging by selecting patients who require neoadjuvant therapy (16).…”

Section: Discussionmentioning

confidence: 99%

Lymph node micrometastases detected by carcinoembryonic antigen mRNA affect long-term survival and disease-free interval in early-stage lung cancer patients

et al. 2012

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Abstract. The majority of stage I lung cancer patients undergo a complete resection of their tumor; however, they still harbor a considerable risk of mortality due to recurrences. A correlation between the presence of lymph node micrometastases and poor prognosis has been observed. The aim of this study was to correlate the lymph node molecular staging with the 5-year survival and disease-free interval following pulmonary lobectomy for non-small cell lung cancer (NSCLC). A quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for carcinoembryonic antigen (CEA) mRNA was performed on primary lung tumors and regional lymph nodes from 55 surgically resected NSCLC patients classified as clinical stage I. CEA mRNA was found to be present in all the primary tumors. RT-PCR revealed the presence of cancer cells in the lymph nodes of 20 patients (36.3%) and routine staining detected lymph node metastases in 11 patients. Significant differences in survival and disease-free intervals were observed in patients with lymph node micrometastases versus patients with negative lymph nodes (P=0.0026 and P=0.0044, respectively). Multivariate analyses confirmed that micrometastases were an independent predictor for worse prognosis (P=0.0098) and a short disease-free interval (P=0.0137). This study demonstrated strong correlations between the molecular detection of lymph node micrometastases and 5-year survival rates and disease-free interval in patients who underwent pulmonary lobectomy for early-stage lung cancer.

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“…According to the patterns on agarose gel electrophoresis and turbidity detection by naked eyes, the best amplification results were obtained with the reaction composing of 0.8 mM dNTPs, 0.7 M bentaine and 6 mM MgSO 4 and performing at 63 °C for 60 min (Fig 2 and 3). Therefore, the DNAs from nine N. bombycis isolates were conducted under this optimal condition and all positive results were presented as shown in Fig 4. Several previous studies reported that the optimal concentration of dNTPs ranged between 0.8 -1.4 mM (Curtis et al, 2008;Maeda et al, 2009;Wei et al, 2014). Among all factors analyzed, MgSO 4 had the greater effect on LAMP reaction since free Mg 2+ affects DNA polymerase activity, primer annealing and magnesium pyrophosphate formation (Mori et al, 2001;Yeh et al, 2005).…”

Section: Lamp Reaction Optimizationmentioning

confidence: 98%

Evaluation of combined effect of micronutrients and rhizobial inoculation on mungbean productivity

Kampliw

Monthatong

2019

IJARe

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In present study, Loop Mediated Isothermal Amplification (LAMP) assay was conducted for diagnosis Nosema bombycis, the pebrine disease pathogen in domestic silkworm, Bombyx mori. Nine isolates of N. bombycis were collected from infected silkworms in rearing areas in Khon Kaen province, Thailand. N. bombycis genomic DNAs were extracted by boiling method and used as templates in LAMP and PCR reactions. A LAMP primer set was designed specific to N. bombycis small subunit ribosomal RNA (SSU rRNA) gene. The results revealed that the optimal condition was constantly performed at 63oC for 1 hour. The product was directly visualized by naked eye and confirmed with agarose gel eletrophoresis. LAMP assay is more sensitive than traditional PCR, since LAMP was able to detect the least 10 spores/ml while PCR needs 100 spores/ml. In addition, the present novel LAMP primer set was specific only to N. bombycis proven by the negative results when other B. mori pathogen DNAs were tested. In conclusion, the LAMP assay demonstrated a great potential alternative method in diagnosis N. bombycis with high sensitivity, rapidity and accuracy which can apply for pebrine disease surveillance.

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Rapid diagnosis of lymph node metastasis in lung cancer with loop-mediated isothermal amplification assay using carcinoembryonic antigen–mRNA

Cited by 12 publications

References 23 publications

Rapid diagnosis of micrometastasis of gastric cancer using reverse transcription loop-mediated isothermal amplification

Rapid diagnosis of micrometastasis of gastric cancer using reverse transcription loop-mediated isothermal amplification

Lymph node micrometastases detected by carcinoembryonic antigen mRNA affect long-term survival and disease-free interval in early-stage lung cancer patients

Evaluation of combined effect of micronutrients and rhizobial inoculation on mungbean productivity

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