The value of three proteins in diagnosing Mycobacterium tuberculosis infection

doi:10.19982/j.issn.1000-6621.20240120

Abstract

Abstract:

Objective: To investigate the diagnostic efficacy of three proteins—Krüppel-like transcription factor 2 (KLF2), guanylate-binding protein 5 (GBP5), and dual specificity phosphatase 3 (DUSP3)—and their assorted combinations in the detection of Mycobacterium tuberculosis (MTB) infection. Methods: Between January and March 2023, patients from Beijing Chest Hospital, Capital Medical University, and individuals undergoing concurrent health examinations were enrolled as study participants. Based on established tuberculosis diagnostic criteria, participants were categorized into three groups: active tuberculosis (ATB) group, consisting of 145 cases; latent tuberculosis infection (LTBI) group, also comprising 145 cases; and a healthy control (HC) group, which included 200 cases. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify the serum levels of KLF2, GBP5, and DUSP3 at 450 nm wavelength absorbance, with results reported as median (interquartile range). The diagnostic potential of each protein, individually and in combination, was analyzed for detecting MTB infection using subject working characteristic curves. Results: Serum levels of KLF2 in the HC group were 0.317 (0.198, 0.496), significantly higher than those in the ATB group at 0.234 (0.160, 0.297) and in the LTBI group at 0.224 (0.145, 0.320), demonstrating statistical significance (P<0.001). Similarly, GBP5 levels in the ATB group were 0.327 (0.259, 0.402), significantly higher than those in the LTBI group at 0.196 (0.150, 0.273) and in the HC group at 0.180 (0.125, 0.281), with statistical significance (P<0.001). The serum level of DUSP3 in the ATB group was 0.329 (0.223, 0.458), significantly exceeding those in the LTBI group at 0.213 (0.160, 0.248) and in the HC group at 0.196 (0.132, 0.297), with statistical significance (P<0.001). Furthermore, the diagnostic performance of the dual protein combination of GBP5 and DUSP3 in differentiating ATB from LTBI, with an area under the curve (AUC) of 0.800, was found to be superior to that of single proteins and their various combinations, such as KLF2 (AUC of 0.534), GBP5 (AUC of 0.761), and DUSP3 alone (AUC of 0.720). For the dual protein combination of GBP5 and DUSP3 in differentiating ATB from LTBI, the sensitivity at the maximum Youden index reached 73.79%, with a specificity of 75.86%. This combination demonstrated superior diagnostic efficacy (AUC of 0.800) compared to the individual proteins and their various combinations. When distinguishing ATB from HC, this dual protein combination also excelled, achieving an AUC of 0.781, with sensitivity at the maximum Youden index at 77.93% and specificity at 71.50%. This was more effective than the individual performances of KLF2 (AUC of 0.629), GBP5 (AUC of 0.740), and DUSP3 (AUC of 0.716), and their different combinations. However, the diagnostic efficacy of the three proteins and their combinations in distinguishing LTBI from HC was less effective, with all AUC values being below 0.700. Conclusion: The findings of this study indicate that there is potential for developing immunodiagnostic methods based on protein levels for detecting MTB infection. Utilizing combinations of proteins such as KLF2, GBP5, and DUSP3 could enhance diagnostic accuracy, offering a promising approach to improve the efficacy of TB diagnostics.

Key words: Mycobacterium tuberculosis, Infection, Immunoassay, Diagnosis, differential

CLC Number:

Zhang Muli, Sun Zhaogang, Cao Tingming, Xie Zhongyao. The value of three proteins in diagnosing Mycobacterium tuberculosis infection[J]. Chinese Journal of Antituberculosis, 2024, 46(7): 808-814. doi: 10.19982/j.issn.1000-6621.20240120

Figures/Tables 4

References 30

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组别	人数 (例/名)	年龄 [岁,M(Q₁,Q₃)]	性别[例/名(构成比,%)]
组别	人数 (例/名)	年龄 [岁,M(Q₁,Q₃)]	男性	女性
ATB组	145	45(31,58)	74(51.03)	71(48.97)
LTBI组	145	42(31,57)	72(49.66)	73(50.34)
HC组	200	32(23,49)	82(41.00)	118(59.00)
统计检验值		χ²=31.554	χ²=4.210
P值		<0.001	0.122

组别	KLF2	GBP5	DUSP3
HC组	0.317(0.198,0.496)	0.180(0.125,0.281)	0.196(0.132,0.297)
LTBI组	0.224(0.145,0.320)	0.196(0.150,0.273)	0.213(0.160,0.248)
ATB组	0.234(0.160,0.297)	0.327(0.259,0.402)	0.329(0.223,0.458)
χ²值	27.927	76.299	58.547
P值	<0.001	<0.001	<0.001

蛋白类型分组	≥40岁者	<40岁者	χ²值	P值
KLF2
HC组	0.320(0.145,0.496)	0.306(0.207,0.493)	0.070	0.7909
LTBI组	0.224(0.116,0.315)^a	0.223(0.165,0.343)^a	0.465	0.4952
ATB组	0.246(0.179,0.297)^a	0.194(0.145,0.303)^a	2.624	0.1053
GBP5
HC组	0.193(0.139,0.278)	0.172(0.109,0.286)	2.518	0.1126
LTBI组	0.194(0.144,0.273)	0.196(0.150,0.292)	0.106	0.7453
ATB组	0.324(0.267,0.423)^ab	0.328(0.255,0.390)^ab	0.1772	0.6738
DUSP3
HC组	0.216(0.152,0.289)	0.190(0.120,0.303)	2.530	0.1117
LTBI组	0.214(0.167,0.259)	0.200(0.141,0.240)	0.789	0.3745
ATB组	0.339(0.245,0.491)^ab	0.316(0.187,0.417)^ab	3.230	0.0723

蛋白及组合	AUC(95%CI)值	敏感度 (%)	特异度 (%)	约登指数	Cut-off值	P值
区分ATB和LTBI
KLF2	0.534(0.467~0.600)	91.72	17.24	0.100	>0.106^a	0.319
GBP5	0.761(0.704~0.817)	78.62	68.28	0.469	>0.250^a	<0.001
DUSP3	0.720(0.659~0.781)	68.97	75.17	0.441	>0.249^a	<0.001
KLF2联合GBP5	0.760(0.703~0.816)	78.62	68.28	0.469	>0.457^b	<0.001
KLF2联合DUSP3	0.718(0.658~0.779)	69.66	73.10	0.428	>0.452^b	<0.001
GBP5联合DUSP3	0.800(0.744~0.847)	73.79	75.86	0.500	>0.452^b	<0.001
KLF2、GBP5、DUSP3联合	0.800(0.745~0.848)	78.62	73.10	0.517	>0.427^b	<0.001
区分ATB和HC
KLF2	0.629(0.572~0.688)	75.17	54.00	0.292	<0.299^a	<0.001
GBP5	0.740(0.688~0.793)	78.62	67.50	0.461	>0.249^a	<0.001
DUSP3	0.716(0.661~0.772)	73.10	66.00	0.391	>0.237^a	<0.001
KLF2联合GBP5	0.750(0.697~0.801)	66.90	76.50	0.434	>0.453^b	<0.001
KLF2联合DUSP3	0.749(0.697~0.801)	57.93	78.00	0.389	>0.454^b	<0.001
GBP5联合DUSP3	0.781(0.729~0.826)	77.93	71.50	0.494	>0.374^b	<0.001
KLF2、GBP5、DUSP3联合	0.780(0.733~0.829)	74.48	72.50	0.469	>0.396^b	<0.001
区分LTBI和HC
KLF2	0.658(0.589~0.716)	71.34	57.00	0.273	<0.285^a	<0.001
GBP5	0.542(0.481~0.602)	78.62	35.00	0.136	>0.144^a	0.188
DUSP3	0.532(0.471~0.593)	90.34	22.00	0.123	>0.125^a	0.312
KLF2联合GBP5	0.652(0.594~0.710)	72.41	55.00	0.274	>0.433^b	<0.001
KLF2联合DUSP3	0.655(0.597~0.713)	68.97	59.00	0.280	>0.442^b	<0.001
GBP5联合DUSP3	0.514(0.452~0.575)	50.23	56.50	0.068	<0.421^b	0.669
KLF2、GBP5、DUSP3联合	0.657(0.599~0.715)	71.03	56.00	0.271	>0.435^b	<0.001