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Virchows Archiv

, Volume 466, Issue 2, pp 169–176 | Cite as

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

  • Xiaopeng Hao
  • Yi LiuEmail author
  • Xiru Li
  • Hua Kang
  • Xiang Qu
  • Jianmiao He
  • Haixu Hu
  • Yan Huang
  • Bing Liu
  • Chengze YuEmail author
Original Article
  • 479 Downloads

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.

Keywords

Breast cancer Sentinel lymph node (SLN) Intra-operative diagnostic method Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) 

Notes

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.

Supplementary material

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Xiaopeng Hao
    • 1
  • Yi Liu
    • 2
    Email author
  • Xiru Li
    • 3
  • Hua Kang
    • 4
  • Xiang Qu
    • 5
  • Jianmiao He
    • 6
  • Haixu Hu
    • 2
  • Yan Huang
    • 1
  • Bing Liu
    • 2
  • Chengze Yu
    • 1
    Email author
  1. 1.Department of Breast SurgeryAffiliated Hospital of Academy of Military Medical SciencesBeijingChina
  2. 2.Translational Medicine Center, Laboratory of OncologyAffiliated Hospital of Academy of Military Medical SciencesBeijingChina
  3. 3.Department of General SurgeryGeneral Hospital of People’s Liberation ArmyBeijingChina
  4. 4.Department of General Surgery, Xuanwu HospitalCapital Medical UniversityBeijingChina
  5. 5.Department of General Surgery, Friendship HospitalCapital Medical UniversityBeijingChina
  6. 6.Department of General Surgery309 Hospital of People’s Liberation ArmyBeijingChina

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