The Research Of Reverse Transcriptase-loop Mediated Isothermal Amplifying CK-19 Judge Breast Cancer Sentinel Lymph Node Metastasis

The axillary lymph node condition of breast cancer patient is a vital treatment and prognosis indicator,the clearation of axillary lymph node is considered standard procedure of breast cancer's surgical treatment。Present research reveals that sentinel lymph node biopsy can substitute axillary lymph node dissection,if the sentinel lymph node is positive,the axillary lymph node is cleared ,so the examination of sentinel lymph node is very important,the purpose of this experiment is to establish a rapid,accurate and objective examination method of judgeing breast cancer sentinel lymph node metastasis,helping instruct the clinical treatment and improve the quality of life。Method:1,Get the sentinel lymph node of 30 early breast cancer patients by the method of methylene blue,divide the lymph node into four parts(a,b,c,d) equally,a and c are used for conventional pathology examination,b and d are marked and frozed under the condition of -80℃。2,Culture the cell of breast cancer cell line and lymphoma cell line,detect the variability of CK-19 expession by LAMP。3,Add the reverse transcriptase at the base of LAMP,detcet the lymph node which pathology proves positive,verify the possibility of this way。4,Improve the method of experiment,shorten the time of reaction,batch confirmation。Result:1,There are 76 sentinel lymph nodes detected from thirty patients,which the positive is 23,the average number is 2.53(76/30), the positive incidence is 30.3%(23/76)。2,The breast cancer which origins from epidermis specifically express CK-19,the detection of CK-19 from normal lymph node means breast cancer metastasis。3,CK-19 can be detected from 3 positive lymph nodes by RT-LAMP,meaning tumor metastasis,the result accords with pathological section HE chromoscopy examination。4,The total reaction can be finished in 1 hour by improving experiment method,this experiment reveals that the positive incidence of sentinel lymph node which is detected by RT-LAMP is 44.7%, increasing 14.4% in contrast with pathological section HE chromoscopy examination's 30.3%。Conclusion:The reaction of RT-LAMP detecting CK-19 can rapidly,accurately and objectively judge the condition of sentinel lymph node,showing favourable application prospect。...