Diagnostic value of combining HE staining with three pathogen detection methods in tuberculosis histopathology for improving the positivity rate of active tuberculosis

doi:10.19983/j.issn.2096-8493.20260072

Abstract

Abstract:

Objective: To explore the clinical value of hematoxylin-eosin (HE) staining combined with acid-fast bacilli (AFB) smear microscopy, Mycobacterium tuberculosis DNA (TB-DNA) detection and GeneXpert MTB/RIF (Xpert) assay in improving the positive detection rate of active tuberculosis in pathological tissues. Methods: Following the inclusion criteria, clinical data from 186 pulmonary tuberculosis patients admitted to Shenzhen Third People’s Hospital in 2023 were collected. The data included postoperative lesion pathological HE staining, AFB smear microscopy, TB-DNA and Xpert results. Patients were classified into four groups based on the inflammatory necrosis progression of postoperative lesions:granulomatous necrosis group (33 cases), general necrosis group (44 cases), caseous necrosis group (81 cases), and caseating necrosis group (28 cases). By comparing the positivity rates of histopathological HE staining with those of AFB smear microscopy, TB-DNA testing, and Xpert testing, we evaluated the diagnostic value of combining conventional histopathological HE staining with these three pathogen detection methods in improving the detection rate of active tuberculosis. Results: The positive rates of Xpert and TB-DNA (77.96% (145/186) and 64.52% (120/186)) were significantly higher than that of AFB staining (46.24% (86/186)) across all groups (χ²=39.757, P<0.001; χ²=12.575, P<0.001). Xpert had a higher overall positive rate than TB-DNA (χ²=8.199, P=0.004). Xpert achieved the highest positivity rate (83.95% (68/81)) in the caseous necrosis group. Whereas the granulomatous inflammation group showed the lowest positivity rate (66.67% (22/33)). The overall positive rates of HE+Xpert (87.10% (162/186)) and the four-method combination (93.01% (173/186)) were significantly higher than that of HE+AFB staining (76.88% (143/186))(χ²=6.571, P=0.010; χ²=18.919, P<0.001). Subgroup analysis revealed that single HE staining consistently yielded the low positive rates across all pathological subtypes:granulomatous inflammation (45.45%, 15/33), general necrosis (77.27%, 34/44), caseous necrosis (86.42%, 70/81), and coagulative necrosis (85.71%, 24/28). The positive rates of HE+Xpert were 75.76% (25/33), 86.36% (38/44), 91.36% (74/81), and 89.29% (25/28). Conclusion: Accurate tissue sampling combined with multi-method detection markedly improves the diagnostic positivity of tuberculous pathological testing. Routine incorporation of molecular techniques such as Xpert significantly enhance the diagnostic positive rate. This is particularly beneficial for detecting granulomatous inflammation, providing crucial evidence for clinical diagnosis.

Key words: Tuberculosis, Biopsy, Histocytochemistry, Granulation tissue, Molecular diagnostic techniques

CLC Number:

R52
R446.8

Zhang Yaxi, Yu Hong, Li Jing, Gou Jizhou, Yang Qianting, Zhan Senlin, Chen Qi. Diagnostic value of combining HE staining with three pathogen detection methods in tuberculosis histopathology for improving the positivity rate of active tuberculosis[J]. Journal of Tuberculosis and Lung Disease , 2026, 7(4): 487-492. doi: 10.19983/j.issn.2096-8493.20260072

Figures/Tables 2

References 19

[1]	屈燕, 李涛, 马文斌, 等. 世界卫生组织《2025年全球结核病报告》解读[J]. 结核与肺部疾病杂志, 2025, 6(6):613-623. doi:10.19983/j.issn.2096-8493.20250178.
[2]	Li T, Yan X, Du X, et al. Extrapulmonary tuberculosis in China:a national survey[J]. Int J Infect Dis, 2023, 128:69-77. doi:10.1016/j.ijid.2022.12.005.
[3]	Dai L, Peng L, Fang T, et al. Diagnostic performance analysis of Xpert MTB/RIF in lymph node tuberculosis:A retrospective study[J]. Diagn Micr Infec Dis, 2024, 110(1):116385. doi:10.1016/j.diagmicrobio.2024.116385.
[4]	Liu J, Qian H, Jin J, et al. Use of metagenomic next-generation sequencing for accurate diagnosis of tuberculous pleurisy:a retrospective cohort study[J]. J Thorac Dis, 2025, 17(9):6771-6778. doi:10.21037/jtd-2024-2050.
[5]	Rao A K, Shastri U D. Role of Bronchoscopic Cytology in Diagnosis of Pulmonary Lesions[J]. J Cytol, 2023, 40(1):35-41. doi:10.4103/joc.joc_138_22. pmid: 37179959
[6]	杨松, 郭建琼, 李同心, 等. 结核病介入诊断与治疗年度进展2023[J]. 中华结核和呼吸杂志, 2024, 47(4):363-370. doi:10.3760/cma.j.cn112147-20231114-00308.
[7]	中华医学会结核病学分会, 结核病病理学诊断专家共识编写组. 中国结核病病理学诊断专家共识[J]. 中华结核和呼吸杂志, 2017, 40(6):419-425. doi:10.3760/cma.j.issn.1001-0939.2017.06.005.
[8]	中华人民共和国国家卫生和计划生育委员会. WS 288-2017肺结核诊断[J]. 结核与肺部疾病杂志, 2024, 5(4):376-378. doi:10.19983/j.issn.2096-8493.2024022.
[9]	Naidoo K, Perumal R, Cox H, et al. The epidemiology, transmission, diagnosis, and management of drug-resistant tuberculosis-lessons from the South African experience[J]. Lancet Infect Dis, 2024, 24(9):e559-e575. doi:10.1016/S1473-3099(24)00144-0. pmid: 38527475
[10]	Iqbal A, Shafique M, Zahoor M A, et al. The occurrence of multidrug-resistant Mycobacterium tuberculosis from patients of pulmonary tuberculosis[J]. J Infect Dev Ctries, 2022, 16(4):698-704. doi:10.3855/jidc.14990.
[11]	王胜芬, 赵雁林. 升级结核病防治策略,提高结核病患者发现水平[J]. 实用医学杂志, 2023, 39(11):1331-1334. doi:10.3969/j.issn.1006-5725.2023.11.002.
[12]	Sun W, Dong Z, Zhou Y, et al. Early Identification of Fungal and Mycobacterium Infections in Pulmonary Granulomas Using Metagenomic Next-Generation Sequencing on Formalin fixation and paraffin embedding tissue[J]. Expert Rev Mol Diagn, 2022, 22(4):461-468. doi:10.1080/14737159.2022.2052046.
[13]	Sun W W, Dong Z W, Zhou Y M, et al. Improving the identification and diagnostic efficiency of Metagenomic Next-Generation Sequencing for mycobacterial granuloma on postoperative formalin-fixed paraffin-embedded specimens[J]. Microbes Infect, 2023, 25(8):105185. doi:10.1016/j.micinf.2023.105185.
[14]	向双, 杨杰斌, 沈昌琼, 等. GeneXpert MTB/RIF检测在石蜡组织结核病理诊断中的应用价值[J]. 临床与实验病理学杂志, 2022, 38(8):1005-1007. doi:10.13315/j.cnki.cjcep.2022.08.025.
[15]	Liu Q, Xu F, Liu Q, et al. Comparative analysis of fiveetiological detecting techniques for the positive rates in the diagnosis of tuberculous granuloma[J]. J Clin Tuberc Other Mycobact Dis, 2023, 32:100378. doi:10.1016/j.jctube.2023.100378.
[16]	Araújo R M, Montenegro R A, Peixoto A D S, et al. Performance of IS6110-LAMP assay for detection of Mycobacterium tuberculosis complex in blood and urine samples from patients with extrapulmonary tuberculosis[J]. Tuberculosis (Edinb), 2023, 143:102423. doi:10.1016/j.tube.2023.102423.
[17]	云海龙, 罗艳, 廖丹丹, 等. 基于石蜡包埋组织比较荧光PCR方法和抗酸染色法对结核分枝杆菌的检出率[J]. 诊断病理学杂志, 2023, 30(7):712, 716. doi:10.3969/j.issn.1007-8096.2023.07.028.
[18]	Lo C K, Purych D, Sekirov I, et al. Evaluation of GeneXpert MTB/Rif Ultra assay performance on formalin-fixed paraffin-embedded tissues for Mycobacterium tuberculosis detection[J]. J Med Microbiol, 2024, 73(10):001918. doi:10.1099/jmm.0.001918.
[19]	Du W L, Song J, Wang J G, et al. Performance of Xpert MTB/RIF Ultra and Xpert MTB/RIF for the Diagnosis of Tuberculosis through Testing of Formalin-fixed Paraffin-embedded Tissues[J]. Biomed Environ Sci, 2019, 32(12):922-925. doi:10.3967/bes2019.115. pmid: 31918797

方法	肉芽肿性炎组(33例)	一般性坏死组(44例)	干酪样坏死组(81例)	凝固性坏死组(28例)	合计(186例)
AFB	6(18.18)	20(45.45)	46(56.79)	14(50.00)	86(46.24)
Xpert	22(66.67)	34(77.27)	68(83.95)	21(75.00)	145(77.96)
χ²值	15.880	9.394	14.329	3.733	39.757
P值^a	<0.001	0.002	<0.001	0.053	<0.001
TB-DNA	21(63.63)	29(65.91)	53(65.43)	17(60.71)	120(64.52)
χ²值	14.102	3.730	1.273	0.650	12.575
P值^b	<0.001	0.053	0.259	0.420	<0.001
χ²值	0.067	1.397	1.272	1.310	8.199
P值^c	0.796	0.237	0.259	0.252	0.004
χ²值	19.477	9.802	14.513	3.736	40.412
P值^d	<0.001	<0.001	0.004	0.155	<0.001

方法	肉芽肿性炎组(33例)	一般性坏死组(44例)	干酪样坏死组(81例)	凝固性坏死组(28例)	合计(186例)
HE	11(33.33)	30(68.18)	68(83.95)	23(82.14)	132(70.97)
HE+AFB	15(45.45)	34(77.27)	70(86.42)	24(85.71)	143(76.88)
χ²值	1.015	0.916	0.195	0.132	1.687
P值^a	0.313	0.338	0.658	0.716	0.193
HE+Xpert	25(75.76)	38(86.36)	74(91.36)	25(89.29)	162(87.10)
χ²值	6.346	1.222	1.000	0.163	6.571
P值^b	0.011	0.268	0.317	0.686	0.010
HE+TB-DNA	21(63.64)	35(79.55)	72(88.89)	24(85.71)	152(81.72)
χ²值	2.200	0.067	0.228	0.132	1.326
P值^c	0.138	0.795	0.632	0.716	0.249
4种方法	30(90.91)	40(90.91)	77(95.06)	26(92.86)	173(93.01)
χ²值	15.714	3.057	3.600	0.746	18.919
P值^d	<0.001	0.080	0.057	0.387	<0.001