血液来源的新型生物标志物对活动性肺结核的诊断价值初探

doi:10.19982/j.issn.1000-6621.20250380

摘要/Abstract

摘要：

目的: 评估外周血结核病特异性生物标志物对活动性肺结核的诊断价值。方法: 采用前瞻性研究方法,采集2024年6—7月上海市肺科医院结核科收治的10例活动性肺结核患者(结核测序组)和10名健康者(健康测序组)的外周血进行转录组测序,筛选出具有潜在诊断价值的靶基因;再于2025年2—10月分别采集69例活动性肺结核患者(结核验证组)、22例非肺结核患者(非结核验证组)和58名健康者(健康验证组)外周血,采用定量逆转录聚合酶链式反应(qRT-PCR)检测外周血中候选基因的mRNA表达水平,以最终诊断为参照标准,通过敏感度、特异度及受试者工作特征(ROC)曲线下面积(AUC)评估筛选出的生物标志物对活动性肺结核的诊断价值。结果: 通过外周血转录组测序筛选出16个候选基因,经qRT-PCR验证,筛选出3个表达水平升高的有诊断价值的标志物,即TRIM22、FKBP9和ERRFI1;与健康验证组比较,3个标志物在结核验证组患者外周血中的转录水平均明显升高(TRIM22:42.32和14.53、FKBP9:6.41和0.20、ERRRFI1:5.70和0.44),差异均有统计学意义(Z值分别为-5.030、-4.425、-6.044,P值均<0.05)。ROC曲线分析显示,与健康验证组比较,TRIM22对结核验证组检测的敏感度、特异度、AUC值分别为80.00%、91.38%、0.956,FKBP9分别为98.44%、91.67%、0.769,ERRFI1分别为94.29%、97.76%、0.819,但3个基因联合使用的诊断效能无明显提升,AUC值仅为0.796;与非结核验证组患者比较,仅ERRFI1对鉴别肺结核有诊断价值,敏感度、特异度、AUC值分别为87.76%、98.28%、0.972。结论: 活动性肺结核患者外周血中TRIM22、FKBP9和ERRFI1可有效区分活动性肺结核患者与健康人群,具有较好的诊断潜力,有望成为诊断活动性肺结核的新型靶标。

关键词: 结核,肺, 生物学标记, 转录组测序, 诊断

Abstract:

Objective: To evaluate the diagnostic value of tuberculosis-specific biomarkers in peripheral blood for active pulmonary tuberculosis (PTB). Methods: In a prospective study, peripheral blood samples were collected from 10 patients with active PTB (PTB sequencing group) and 10 healthy individuals (control sequencing group) admitted to the Tuberculosis Department of Shanghai Pulmonary Hospital between June and July 2024 for transcriptome sequencing to screen for potential diagnostic target genes. Subsequently, from February to October 2025, peripheral blood samples were collected from 69 PTB patients (TB validation group), 22 non-TB patients (non-TB validation group), and 58 healthy individuals (control validation group) was used to measure. The mRNA expression levels of candidate genes in peripheral blood were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The diagnostic value of the identified biomarkers for active PTB was assessed by calculating sensitivity, specificity, and the area under the receiver operating characteristic curve (AUC) against the final clinical diagnosis reference standard. Results: Sixteen candidate genes were identified by transcriptomic sequencing of peripheral blood. After qRT-PCR validation, three biomarkers with upregulated expression levels were identified: TRIM22, FKBP9, and ERRFI1. Compared to the control validation group, the expression levels of all three markers were significantly higher in the TB validation group (TRIM22: 42.32 vs. 14.53; FKBP9: 6.41 vs. 0.20; ERRFI1: 5.70 vs. 0.44), with statistically significant differences (Z=-5.030, -4.425, -6.044, respectively; all P<0.05). Compared to the control validation group, ROC curve analysis revealed that TRIM22 demonstrated a sensitivity, specificity, and AUC of 80.00%, 91.38%, and 0.956, respectively; FKBP9 had 98.44%, 91.67%, and 0.769, respectively; and ERRFI1 had 94.29%, 97.76%, and 0.819. The diagnostic performance did not improve significantly when the three genes were used in combination (AUC=0.796). When compared to non-TB patients, only ERRFI1 showed diagnostic value in distinguishing active PTB, with a sensitivity of 87.76%, specificity of 98.28%, and an AUC of 0.972. Conclusion: The peripheral blood levels of TRIM22, FKBP9, and ERRFI1 in patients with active PTB can effectively distinguish them from healthy individuals, showing promising diagnostic potential. These markers may serve as novel targets for the diagnosis of active PTB.

Key words: Tuberculosis, pulmonary, Biological markers, Bulk transcriptome sequencing, Diagnosis

中图分类号:

张宇洁, 周洁, 王芸, 范琳. 血液来源的新型生物标志物对活动性肺结核的诊断价值初探[J]. 中国防痨杂志, 2026, 48(3): 320-328. doi: 10.19982/j.issn.1000-6621.20250380

Zhang Yujie, Zhou Jie, Wang Yun, Fan Lin. Evaluation of diagnostic value of novel blood-derived biomarkers for active pulmonary tuberculosis[J]. Chinese Journal of Antituberculosis, 2026, 48(3): 320-328. doi: 10.19982/j.issn.1000-6621.20250380

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参考文献 31

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基因(人)	前引物	后引物
β-actin	5'-CCTGGACTTCGAGCAAGAGATGG-3'	5'-CAGGAAGGAAGGCTGGAAGAGTG-3'
ETV5	5'-TCAGCAAGTCCCTTTTATGGTC-3'	5'-GCTCTTCAGAATCGTGAGCCA-3'
CDKL3	5'-TTCATCACGAAAACCTGGTCAA-3'	5'-AAGTCGCTTACTCTCTAGTCCAT-3'
AJUBA	5'-ATGGGGAAGTCCTATCATCCAG-3'	5'-TGGTAGTCGGTGACACAGTAT-3'
P4HA2	5'-CAAACTGGTGAAGCGGCTAAA-3'	5'-GCACAGAGAGGTTGGCGATA-3'
CFB	5'-GCGATCTGTGACAACGGAG-3'	5'-GCTGTCTTCAAGGCGGTACT-3'
LIPT2	5'-CTGACGCCCGAGGAAACTG-3'	5'-GTGGGATGTGATGTGCCTTC-3'
TUBB3	5'-GGCCAAGGGTCACTACACG-3'	5'-GCAGTCGCAGTTTTCACACTC-3'
BEND7	5'-CTGGATCAAACTGCTGTACTTG-3'	5'-GGGAAATTGGAGGTTGTTGTCTG-3'
DDAH1	5'-CAAAAGGACAAATCAACGAGGT-3'	5'-TGTGCAGATTCACTAGACCCAA-3'
KLHL23	5'-ATGCCACGGAAAGGATATACCT-3'	5'-CAACACCTGAGGACCAAACTAC-3'
MOK	5'-CCGGAGTGTCTCCTCACTGAT-3'	5'-CAGACTGGCGATCTCGTAGAA-3'
DOCK6	5'-GCTTCTGGAGACGAGAGGTC-3'	5'-AGGTTCCTCAGGTCGAAGATG-3'
FKBP9	5'-CAGGTGTCTGATTTTGTGAGGT-3'	5'-TTCATGCGATTGTGACTCGAA-3'
TG	5'-AGGGAGAGTTTATGCCTGTCC-3'	5'-CAATACCCAGATACCTCAGGGA-3'
ERRFI1	5'-GACCCACCGAAGATTAAGAAGG-3'	5'-GGTCTAGGAGGTATGGGAACTCT-3'
TRIM22	5'CTGTCCTGTGTGTCAGACCAG-3'	5'TGTGGGCTCATCTTGACCTCT-3'

基因	健康验证组 (58例)	结核验证组		Z值	P值
基因	健康验证组 (58例)	病原学阳性(35例)	病原学阴性(34例)	Z值	P值
ETV5	0.87(0.48,2.65)	0.18(0.14,0.29)	0.11(0.09,0.47)	4.543	0.077
CDKL3	0.94(0.50,3.09)	0.22(0.17,0.38)	0.36(0.13,0.70)	3.938	0.008
AJUBA	0.40(0.13,1.53)	0.22(0.14,0.35)	0.44(0.29,0.90)	0.379	0.557
P4HA2	0.05(0.02,0.09)	0.02(0.02,0.03)	0.04(0.03,0.06)	3.009	0.002
CFB	0.86(0.35,5.05)	0.29(0.09,0.45)	0.37(0.11,0.56)	2.857	0.323
LIPT2	0.37(0.15,1.03)	0.39(0.32,0.57)	0.40(0.29,0.77)	-0.729	0.647
TUBB3	0.44(0.18,1.42)	0.34(0.18,0.94)	0.45(0.20,1.69)	0.076	0.543
DDAH1	0.30(0.09,1.68)	0.07(0.05,0.10)	0.08(0.05,0.13)	3.304	0.327
KLHL23	1.69(0.79,3.96)	0.02(0.01,0.04)	0.03(0.00,0.07)	6.044	<0.001
BEND7	1.30(0.72,3.72)	6.11(3.86,17.81)	4.36(2.68,12.57)	-4.007	0.911
MOK	0.55(0.21,1.53)	0.31(0.12,0.45)	0.45(0.16,0.63)	1.003	0.752
DOCK6	0.19(0.10,0.35)	0.07(0.05,0.13)	0.08(0.06,0.12)	3.498	0.044
FKBP9	0.20(0.11,0.49)	6.41(4.05,12.32)	5.70(2.54,7.40)	-4.425	0.007
TG	0.27(0.15,0.51)	0.23(0.13,0.36)	0.26(0.15,0.40)	1.065	0.248
ERRGI1	0.44(0.17,1.37)	5.70(3.05,13.46)	5.57(2.95,7.38)	-6.044	0.002
TRIM22	14.53(12.40,15.45)	42.32(29.75,72.16)	43.69(32.85,64.09)	-5.030	<0.001

指标	最佳界值	敏感度(%)	特异度(%)	AUC值
TRIM22
肺结核患者	22.60	80.00	91.38	0.956
病原学阴性	32.33	98.21	93.79	0.896
病原学阳性	32.63	97.21	93.79	0.958
FKBP9
肺结核患者	0.201	98.44	91.67	0.769
病原学阴性	0.206	95.83	86.02	0.731
病原学阳性	3.754	82.35	91.38	0.894
ERRFI1
肺结核患者	0.095	94.29	97.76	0.819
病原学阴性	1.184	85.11	75.86	0.842
病原学阳性	3.542	70.59	94.83	0.838
联合检测
肺结核患者	0.545	88.24	76.11	0.796
病原学阴性	0.685	98.53	97.23	0.848
病原学阳性	0.005	80.39	98.23	0.885

基因	Non-TB组	TB(MTB阳性)组	TB(MTB阴性)组	Z值	P值^a	Z值	P值^b	Z值	P值^c
TRIM22	42.32(30.84,60.16)	42.33(29.76,70.86)	43.15(32.85,88.45)	2.279	0.651	4.507	0.528	1.977	0.778
FKBP9	8.85(3.46,12.42)	5.77(3.84,12.15)	5.70(2.54,7.40)	1.826	0.349	3.362	0.058	1.841	0.085
ERRFI1	0.86(0.23,4.46)	6.41(4.05,12.32)	5.32(2.89,7.47)	1.784	0.001	2.152	0.005	1.672	0.095