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An intra-operative RT-LAMP method allows rapid and reliable detection of sentinel lymph node metastasis in breast cancer patients

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Abstract

The rapid determination of metastasis in sentinel lymph nodes (SLNs) of breast cancer patients plays a significant role in the selection of a surgery strategy. Although a previous one-step nucleic acid amplification assay that uses reverse-transcription (RT) loop-mediated isothermal amplification (LAMP) has showed specific advantages over traditional pathological examination, its target marker requires optimisation. In addition to epithelial-specific CK19, the internal control gene PBGD and the breast-specific PIP were included in the new method. After the RT-LAMP primers were designed and verified using a cell line, the performance of our method was evaluated by comparing it with the corresponding result of the Food and Drug Administration approved breast lymph node (BLN) assay and routine pathological examination. One hundred and seventy-four valid SLN samples from 101 patients were collected from five hospitals. The threshold of reaction time for CK19, PIP and PBGD was defined as 16, 20 and 20 min, respectively. Compared with the BLN assay, the concordance rate of our method was 95.4 % (166/174). Statistical analysis revealed that the two methods are consistent (kappa = 0.890, P < 0.001). When compared with pathological examination, the performance of our method (sensitivity = 81.3 %, specificity = 89.7 %, kappa = 0.691, P < 0.001) was similar to that of the BLN assay (sensitivity = 87.5 %, specificity = 84.9 %, kappa = 0.668, P < 0.001). This result demonstrates the potential usefulness of our method in clinical practice. In conclusion, we preliminarily established an intra-operative diagnostic method that assimilates the merits of previous assays. In contrast with the BLN assay and pathological examination, our method can be completed in 30 min and shows high sensitivity, specificity and consistency, which we consider as promising for clinical application.

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Acknowledgements

We gratefully acknowledge the excellent assistance of Jia Mi and Fengjing Li (from Beijing Lanpu Bio-tech Co., Ltd.) in design of LAMP primer. The study was supported by the Major Special Project of Beijing Municipal Science and Technology Commission (D09050703570905).

Conflict of interest

The authors declare that they have no competing interests.

Author information

Authors and Affiliations

  1. Department of Breast Surgery, Affiliated Hospital of Academy of Military Medical Sciences, No. 8 Dongdajie, Beijing, 100071, China

    Xiaopeng Hao, Yan Huang & Chengze Yu

  2. Translational Medicine Center, Laboratory of Oncology, Affiliated Hospital of Academy of Military Medical Sciences, No. 8 Dongdajie, Beijing, 100071, China

    Yi Liu, Haixu Hu & Bing Liu

  3. Department of General Surgery, General Hospital of People’s Liberation Army, Beijing, China

    Xiru Li

  4. Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing, China

    Hua Kang

  5. Department of General Surgery, Friendship Hospital, Capital Medical University, Beijing, China

    Xiang Qu

  6. Department of General Surgery, 309 Hospital of People’s Liberation Army, Beijing, China

    Jianmiao He

Authors
  1. Xiaopeng Hao
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  2. Yi Liu
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  4. Hua Kang
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  7. Haixu Hu
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  8. Yan Huang
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  9. Bing Liu
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  10. Chengze Yu
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Corresponding authors

Correspondence to Yi Liu or Chengze Yu.

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X. Hao and Y. Liu contributed equally.

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Hao, X., Liu, Y., Li, X. et al. An intra-operative RT-LAMP method allows rapid and reliable detection of sentinel lymph node metastasis in breast cancer patients. Virchows Arch 466, 169–176 (2015). https://doi.org/10.1007/s00428-014-1693-4

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  • DOI: https://doi.org/10.1007/s00428-014-1693-4

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