Application of droplet digital PCR system for detecting NTRK fusions in gastrointestinal adenocarcinomas
10.13315/j.cnki.cjcep.2024.10.009
- VernacularTitle:微滴式数字PCR在检测胃腺癌和结直肠腺癌NTRK基因融合中的应用
- Author:
Binbin LIU
1
,
2
;
Xiaohong PU
;
Yao FU
;
Dongying ZHANG
Author Information
1. 南京中医药大学附属中西医结合医院病理科,南京 210028
2. 江苏省中医药研究院,南京 210028
- Keywords:
gastrointestinal adenocarcinoma;
NTRK fusion;
ddPCR;
FISH;
immunohistochemistry;
NGS
- From:
Chinese Journal of Clinical and Experimental Pathology
2024;40(10):1052-1058
- CountryChina
- Language:Chinese
-
Abstract:
Purpose To evaluate the feasibility of droplet digital PCR(ddPCR)for gene fusion detection of neurotrophic receptor kinase(NTRK).Methods A total of 830 cases of primary colorectal adenocarcinoma(CRAC)and 560 cases of gastric adenocarcinoma(GAC)were retrospectively studied.Im-munohistochemistry(IHC)and fluorescence in situ hybridization(FISH)were used to detect pan-TRK protein expression and NTRK1/2/3 gene fracture in formalin-fixed paraffin-embedded(FFPE)tumor tissues,respectively.FISH or IHC positive sam-ples were further detected by next generation sequencing and ddPCR.Results All FFPE samples were tested successfully by IHC and FISH methods.A total of 26 samples with NTRK gene breaks or pan-TRK expression were detected by IHC,among these 26 cases,21 FISH were positive and 18 IHC positive.A total of 14 cases of NTRK fusion and 2 cases of amplification were detected by DNA sequencing.A total of 3 cases carrying NTRK gene fusion were detected by RNA sequencing,and the results of ddPCR were completely consistent with RNA sequen-cing.Conclusion ddPCR can effectively distinguish false posi-tive CRAC and GAC cases harboring NTRK fusion detected by FISH and DNA sequencing,which can be used as the effective method for screening NTRK gene fusion.
