The potential of 2 antigens of Mycobacterium tuberculosis for the serodiagnosis of latent tuberculosis infection

doi:10.3969/j.issn.1673-8640.2016.08.005

Abstract

Abstract:

Objective To investigate the potential of 2 antigens（Rv3160c and 35kD antigens） of Mycobacterium tuberculosis（MTB） highly expressed during latent infection for the serodiagnosis of latent tuberculosis infection（LTBI）. Methods Rv3160c and 35kD antigens were purified,and the immunogenicity was evaluated by C57BL/6 mouse model. MTB antigens,early secretory antigenic target-6（ESAT-6） and 38kD antigens which were in commercial use,were included as positive controls. Serum specificity antibody reactivities of 20 active pulmonary tuberculosis patients,25 LTBI patients（contacting closely with active pulmonary tuberculosis patients for a long time） and 24 healthy subjects were determined using Rv3160c and 35kD antigens. Serum levels of anti-Rv3160c and anti-35kD antibodies were determined. Results Rv3160c and 35kD antigens induced strong humoral immunity in mice. Serum antibody titre reached 1∶256 000,which was close to that of positive control Ag85A. Serum levels of anti-Rv3160c and anti-35kD antibodies in LTBI group were higher than those in healthy control group （P<0.01）, and serum level of anti-35kD antibody in LTBI group was higher than that in active pulmonary tuberculosis group（P<0.05）. However,serum level of anti-Rv3160c antibody had no statistical significance between LTBI and active pulmonary tuberculosis groups（P>0.05）. Serum levels of anti-ESAT-6 antibody in LTBI and active pulmonary tuberculosis groups were higher than that in healthy control group（P<0.01）,and there was no statistical significance between LTBI and active pulmonary tuberculosis groups（P>0.05）. Serum level of anti-38kD antibody in LTBI group was higher than those in active pulmonary tuberculosis and healthy control groups（P<0.05）,but there was no statistical significance between active pulmonary tuberculosis and healthy control groups（P>0.05）. Both antigens reacted more strongly with sera from LTBI group than sera from healthy control group. The 35kD antigen had a weak reactivity in active pulmonary tuberculosis group,and its pattern of reactivity was similar to that of 38kD antigen. Rv3160c antigen was unable to distinguish active pulmonary tuberculosis and LTBI,and its reactivity pattern was similar to that of ESAT-6 antigen. Conclusions Rv3160c and 35kD antigens are highly specific to LTBI. It is of potential to be used in the diagnosis of LTBI.

Key words: Mycobacterium tuberculosis, Rv3160c antigen, 35kD antigen, Latent infection, Serodiagnosis

CLC Number:

R446.1

SUN Ruifeng, XIANG Zhihao, CHEN Fuzeng, RU Huanwei, MAI Juntao, YUAN Li, LIU Jun. The potential of 2 antigens of Mycobacterium tuberculosis for the serodiagnosis of latent tuberculosis infection[J]. Laboratory Medicine, 2016, 31(8): 656-661.

Figures/Tables 4

References 17

[1]	World Health Organization.Global tuberculosis report 2015[R]. Geneva:WHO,2015.
[2]	LIENARDT C,GLAZIOU P,UPLEKAR M,et al.Global tuberculosis control:lessons learnt and future prospects[J]. Nat Rev Microbiol,2012,10(6):407-416.
[3]	PAI M.Spectrum of latent tuberculosis-existing tests cannot resolve the underlying phenotypes[J]. Nat Rev Microbiol,2010,8(3):242.
[4]	郭建,范齐文,范小勇,等. 结核分枝杆菌rdESAT-6抗原在耻垢分枝杆菌中的制备及其血清学诊断研究[J]. 检验医学,2014,29(6):607-612.
[5]	钟华清,徐梦华,卢丽娟,等. 干扰素-γ释放试验在儿童结核感染中的应用价值[J]. 检验医学,2016,31(3):176-179.
[6]	LOSI M,BOSSINK A,CODECASA L,et al.Use of a T-cell interferon-gamma release assay for the diagnosis of tuberculous pleurisy[J]. Eur Respir J,2007,30(6):1173-1179.
[7]	CHEN JM,REN H,SHAW JE,et al.Lsr2 of Mycobacterium tuberculosis is a DNA-bridging protein[J]. Nucleic Acids Res,2008,36(7):2123-2135.
[8]	LIU J,GORDON BR.Targeting the global regulator Lsr2 as a novel approach for anti-tuberculosis drug development[J]. Expert Rev Anti Infect Ther,2012,10(9):1049-1053.
[9]	GORDON BR,LI Y,WANG L,et al.Lsr2 is a nucleoid-associated protein that targets AT-rich sequences and virulence genes in Mycobacterium tuberculosis[J]. Proc Natl Acad Sci USA,2010,107(11):5154-5159.
[10]	SHANKAR EM,VIGNESH R,ELLEGARD R,et al.HIV-Mycobacterium tuberculosis co-infection:a 'danger-couple model' of disease pathogenesis[J]. Pathog Dis,2014,70(2):110-118.
[11]	VINNARD C,LINKIN D,BEHRMAN A.Current approach to latent tuberculosis diagnosis and treatment among medical center occupational health physicians[J]. Infect Control Hosp Epidemiol,2012,33(12):1262-1265.
[12]	MAZUREK GH,VILLARINO ME,CDC. Guidelines for using the QuantiFERON-TB test for diagnosing latent Mycobacterium tuberculosis infection[J]. MMWR Recomm Rep,2003,52(RR-2):15-18.
[13]	RAVN P,MUNK ME,ANDERSEN B,et al.Prospective evaluation of a whole-blood test using Mycobacterium tuberculosis-specific antigens ESAT-6 and CFP-10 for diagnosis of active tuberculosis[J]. Clin Diagn Lab Immunol,2005,12(4):491-496.
[14]	WAYNE LG,HAYES LG.An in vitro model for sequential study of shiftdown of Mycobacterium tuberculosis through two stages of nonreplicating persistence[J]. Infect immune,1996,64(6):2062-2069.
[15]	CHAO MC,RUBIN EJ.Letting sleeping dos lie:does dormancy play a role in tuberculosis?[J]. Ann Rev Microbiol,2010,64:293-311.
[16]	BALHANA RJ,SINGLA A,SIKDER MH,et al.Global analyses of TetR family transcriptional regulators in mycobacteria indicates conservation across species and diversity in regulated functions[J]. BMC Genomics,2015,16:479.
[17]	PARK HD,GUINN KM,HARRELL MI,et al.Rv3133c/dosR is a transcription factor that mediates the hypoxic response of Mycobacterium tuberculosis[J]. Mol microbiol,2003,48(3):833-843.

引物	序列（5'~3'）
Rv3160c	F：CGTGGATCCATGCCGAGGCAGGCCG- GCCGCTG
	R：GCAAAGCTTCTAGAGCCCGCGGTCG- GGGGGTGCG
Rv2744c	F：GCGCATATGATGGCCAATCCGTTCGT- TAAAGC
	R：TATGTCGACCTACTGACCGTAGGGCT- GCTCGC

引物	序列（5'~3'）
Rv3160c	F：CGTGGATCCATGCCGAGGCAGGCCG- GCCGCTG
	R：GCAAAGCTTCTAGAGCCCGCGGTCG- GGGGGTGCG
Rv2744c	F：GCGCATATGATGGCCAATCCGTTCGT- TAAAGC
	R：TATGTCGACCTACTGACCGTAGGGCT- GCTCGC