Yan Liang

309th Hospital of the PLA, Peping, Beijing, China

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Publications (19)35.11 Total impact

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    ABSTRACT: The latency-associated antigen Rv2659c is a starvation-related protein of Mycobacterium tuberculosis (M. tuberculosis). It has potential use in tuberculosis (TB) control, but its immunological characteristics in Chinese populations are unclear. In this study, immunological characteristics and potential diagnostic use of recombinant Rv2659c protein were assessed. Interferon-γ (IFN-γ) production from peripheral blood mononuclear cells (PBMC) was assayed by enzyme-linked immunospot (ELISPOT) in TB patients (80 cases), individuals who were purified protein derivative (PPD)-positive after Bacillus Calmette-Guérin (BCG) vaccination (27 cases), nontuberculous respiratory disease patients (30 cases), individuals who were identified by standard techniques as having latent TB infection (LTBI) (37 cases), and uninfected healthy individuals (75 cases). Serum immunoglobulin G (IgG) levels were assayed by enzyme-linked immunosorbent assay (ELISA) in TB patients (43 cases), LTBI individuals (36 cases) and uninfected healthy individuals (66 cases). When stimulated by rRv2659c, PBMC from LTBI individuals gave ELISPOT counts that were significantly higher than those from TB patients, BCG vaccinated individuals, non-TB respiratory disease patients and uninfected healthy individuals (p < 0.05). The rRv2659c stimulation gave detectable IFN-γ production in a higher proportion of persons with LTBI compared with TB patients and uninfected healthy individuals. BCG vaccination and non-TB respiratory disease had little influence on the PBMC response to rRv2659c. The levels of serum IgG specific for rRv2659c were not significantly different between LTBI individuals and TB patients (p > 0.05). These results suggest that rRv2659c has potential for the diagnosis of LTBI. This is the first clinical report of human immune recognition of Rv2659c in Chinese populations.
    Journal of microbiology, immunology, and infection = Wei mian yu gan ran za zhi 04/2014; · 1.63 Impact Factor
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    ABSTRACT: Background The latency-associated antigen Rv2659c is a starvation-related protein of Mycobacterium tuberculosis (M. tuberculosis). It has potential use in tuberculosis (TB) control, but its immunological characteristics in Chinese populations are unclear. Methods In this study, immunological characteristics and potential diagnostic use of recombinant Rv2659c protein were assessed. Interferon-γ (IFN-γ) production from peripheral blood mononuclear cells (PBMC) was assayed by enzyme-linked immunospot (ELISPOT) in TB patients (80 cases), individuals who were purified protein derivative (PPD)-positive after Bacillus Calmette-Guérin (BCG) vaccination (27 cases), nontuberculous respiratory disease patients (30 cases), individuals who were identified by standard techniques as having latent TB infection (LTBI) (37 cases), and uninfected healthy individuals (75 cases). Serum immunoglobulin G (IgG) levels were assayed by enzyme-linked immunosorbent assay (ELISA) in TB patients (43 cases), LTBI individuals (36 cases) and uninfected healthy individuals (66 cases). Results When stimulated by rRv2659c, PBMC from LTBI individuals gave ELISPOT counts that were significantly higher than those from TB patients, BCG vaccinated individuals, non-TB respiratory disease patients and uninfected healthy individuals (p < 0.05). The rRv2659c stimulation gave detectable IFN-γ production in a higher proportion of persons with LTBI compared with TB patients and uninfected healthy individuals. BCG vaccination and non-TB respiratory disease had little influence on the PBMC response to rRv2659c. The levels of serum IgG specific for rRv2659c were not significantly different between LTBI individuals and TB patients (p > 0.05). Conclusion These results suggest that rRv2659c has potential for the diagnosis of LTBI. This is the first clinical report of human immune recognition of Rv2659c in Chinese populations.
    Journal of Microbiology, Immunology and Infection. 01/2014;
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    ABSTRACT: Tuberculosis (TB), an infectious disease caused by infection of Mycobacterium tuberculosis, is a major public health challenge globally. Genetic epidemiological evidence suggests a genetic basis for TB, but the molecular mechanism for a genetic predisposition to TB remains largely unknown. Thirty-five tag single-nucleotide polymorphisms (SNPs) across 11 candidate cytokines and related genes, including IL-12/IFN-γ axis genes (IL12B, IL12RB1, IL18R1, IL27, IFNGR1, IFNGR2 and STAT1), the TNF gene locus (TNF and LTA), IL10, and CCL2, were genotyped using Sequenom's iPLEX assays in 1,032 patients with TB and 1,008 controls of Chinese Han origin. We did not find that any of the 35 tag SNPs individually or as haplotypes was significantly associated with susceptibility to TB, on the basis of multivariable logistic regression analysis with adjustment for age and sex. However, stratification analyses showed that, in those with age 46 years or older, carrying the rs1974675 T allele in the IL18R1 gene had a significantly decreased susceptibility to TB occurrence compared with carrying the C/C genotype (OR = 0.57, P = 5.0×10-4). Further analysis indicated that a SNP in absolute linkage disequilibrium with rs1974675, rs3755276, is located within a CpG dinucleotide and showed hypomethylation in controls than in patients (19.6% vs. 31.4%; P = 1.0×10-4) and genotype-specific DNA methylation at the IL18R1 promoter and IL18R1 mRNA levels. In addition, DNA methylation levels were significantly inversely correlated with mRNA levels. Thus, decreased mRNA levels of IL18R1 due to rs3755276 may partially mediate the increased susceptibility to TB risk.
    PLoS ONE 01/2014; 9(10):e110734. · 3.53 Impact Factor
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    ABSTRACT: Rapid antibody detection has been used widely as a diagnostic aid in the diagnosis of tuberculosis (TB), especially for bacterium-negative TB. We evaluated the diagnostic value of four commercial antibody detection kits with 501 sera from TB patients and controls. The sensitivities and specificities of the four kits were as follows: (1) Rapid test A 71.8% and 91.4% (85.5% for non-TB diseases and 92.8% for healthy controls); (2) Rapid test B 74.2% and 83.1% (63.9% for non-TB diseases and 92.8% for healthy controls); (3) Rapid test C 21.0% and 95.2% (for non-TB diseases); (4) Rapid test D 6.3% and 98.8% (for non-TB diseases). Overall the four commercial serological antibody detection kits performed very differently. Therefore, careful evaluation should be undertaken before any of these commercial serological assays can be recommended as a screening test for TB.
    Annals of clinical and laboratory science 01/2013; 43(1):101-4. · 0.88 Impact Factor
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    ABSTRACT: The situation of tuberculosis (TB) is very severe in China. New therapeutic agents or regimens to treat TB are urgently needed. In this study, Mycobacterium tuberculosis-infected mice were given immunotherapy intramuscularly with Ag85A/B chimeric DNA or saline, plasmid vector pVAX1, or Mycobacterium vaccae vaccine. The mice treated with Ag85A/B chimeric DNA showed significantly higher numbers of T cells secreting interferon-gamma (IFN-γ), more IFN-γ in splenocyte culture supernatant, more Th1 and Tc1 cells, and higher ratios of Th1/Th2 and Tc1/Tc2 cells in whole blood, indicating a predominant Th1 immune response to treatment. Infected mice treated with doses of 100 μg Ag85A/B chimeric DNA had an extended time until death of 50% of the animals that was markedly longer than the saline and vector control groups, and the death rate at 1 month after the last dose was lower than that in the other groups. Compared with the saline group, 100 μg Ag85A/B chimeric DNA and 100 μg Ag85A DNA reduced the pulmonary bacterial loads by 0.79 and 0.45 logs, and the liver bacterial loads by 0.52 and 0.50 logs, respectively. Pathological changes in the lungs were less, and the lesions were more limited. These results show that Ag85A/B chimeric DNA was effective for the treatment of TB, significantly increasing the cellular immune response and inhibiting the growth of M. tuberculosis.
    FEMS Immunology & Medical Microbiology 12/2012; 66(3):419-26. · 2.68 Impact Factor
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    ABSTRACT: To study the relationship between the genetic polymorphisms of carboxylesterase 1 gene (CES1) and the susceptibility to antituberculosis drug-induced hepatotoxicity (ATBDIH). Genetic polymorphisms of CES1 in 473 tuberculosis patients with or without hepatotoxicity (200:273) after antituberculosis chemotherapy were analyzed by PCR-MassArray. In 4 tags of CES1 single nucleotide polymorphism (SNP), the frequency of the rs1968753 allele had statistical difference between the hepatotoxicity group and the no-hepatotoxicity group(P = 0.0236). The characteristics of anti-hepatotoxicity had been shown relationship with rs8192950 (P = 0.044, OR = 0.649, 95%CI = 0.426 - 0.989, AC/AA) and rs1968753 (P = 0.048, OR = 0.556, 95%CI = 0.311 - 0.995, GG/AA). The diplotypes with 'CGC' haplotype exhibited significant protection against hepatotoxicity at one copy (P = 0.048, OR = 0.654, 95%CI = 0.430 - 0.996). The genetic polymorphisms of CES1 might have significant association with ATBDIH. SNP rs8192950 AC genotype and rs1968753 GG genotype might be the candidates for risk prediction of ATBDIH.
    Zhonghua nei ke za zhi [Chinese journal of internal medicine] 07/2012; 51(7):524-30.
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    ABSTRACT: 1. The present study investigated the relationship between antituberculosis (anti-TB) drug-induced hepatotoxicity and genetic polymorphisms of two important drug-metabolizing enzymes involved in the metabolism of isoniazid, namely N-acetyltransferase 2 (NAT2) and cytochrome P450 2E1 (CYP2E1). 2. A polymerase chain reaction direct sequencing approach was used to detect genetic polymorphisms of the NAT2 and CYP2E1 genes in tuberculosis (TB) patients with (n = 101) or without (n = 107) anti-TB drug-induced hepatotoxicity. Associations between various genetic polymorphisms and anti-TB drug-induced hepatotoxicity were then determined. 3. Patients with NAT2 (282TT , 590AA and 857GA) alleles had an increased susceptibility to anti-TB drug-induced hepatotoxicity. The slow acetylator NAT2 genotypes (especially NAT2*6A/7B and NAT2*6A/6A) were risk factors for hepatotoxicity (odds ratio (OR) 9.57 (P < 0.001) for NAT2*6A/7B; OR 5.24 (P = 0.02) for NAT2*6A/6A). 4. The CYP2E1 genotype per se was not significantly associated with the development of anti-TB drug-induced hepatotoxicity. However, the combination of the CYP2E1 C1/C1 genotype with a slow acetylator NAT2 genotype increased the risk of anti-TB drug-induced hepatotoxicity (OR 5.33; P = 0.003) compared with the combination of a rapid acetylator NAT2 genotype with either a C1/C2 or C2/C2 genotype. 5. Thus, slow acetylators with the NAT2*6A/7B and NAT2*6A/6A genotypes combined with the C1/C1 CYP2E1 genotype may be involved in the pathogenesis of anti-TB drug-induced hepatotoxicity. 6. The present findings may be explained, in part, by changes in the metabolism of the anti-TB drug isoniazid induced via NAT2 and CYP2E1, a metabolic process known to produce hepatotoxic intermediates.
    Clinical and Experimental Pharmacology and Physiology 04/2012; 39(6):535-43. · 2.41 Impact Factor
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    ABSTRACT: The diagnosis for smear-negative pulmonary tuberculosis (TB) is very difficult. Proteomic fingerprinting of sera is a potentially useful tool. This study analyzed the results of the proteomic fingerprinting in sera obtained from active TB patients and controls using the surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) and protein-chip technology. The peaks were detected and analyzed, and a diagnostic system was developed. The protein peak was identified using high performance liquid chromatography (HPLC)-tandem matrix-assisted laser desorption/ionization-TOF-MS (MALDI-TOF-MS). Around 50 protein peaks were found significantly different between the TB patients and the controls (P<0.01). Three protein peaks at m/z 5643, 4486 and 4360 were selected for system classification and the development of a decision model. The model distinguished the TB patients from the controls with a sensitivity of 96.9% and a specificity of 97.8%, respectively. The diagnostic accuracy was up to 97.3%. The one most discrepant protein peak at m/z 5643 seen in sera of active TB patients was identified as orosomucoid. A diagnostic system for active TB was developed using mass spectrometry and protein chip technology and required only small-volume serum samples. One potential protein biomarker at m/z 5643 was identified as orosomucoid.
    Clinica chimica acta; international journal of clinical chemistry 02/2012; 413(9-10):883-7. · 2.54 Impact Factor
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    ABSTRACT: China is a country with a high prevalence of latent tuberculosis (TB) infection (LTBI) and has a policy of routine bacillus Calmette-Guérin (BCG) vaccination. The purified protein derivative (PPD) skin test cannot distinguish TB infection from BCG vaccination, and the diagnosis of LTBI lacks an accepted standard method. The primary objective of this study was to assess the potential of a highly sensitive whole-blood interferon (IFN)-γ release assay that uses recombinant culture filtrate protein (CFP)-10/early secretory antigenic target (ESAT)-6 fusion protein (rCFP-10/ESAT-6) as a stimulus for diagnosis of LTBI. Between December 2009 and March 2010, a total of 892 new recruits to the army in Beijing, China, were interviewed and routinely examined by chest radiographs. IFNγ released from whole blood in response to stimulation with rCFP-10/ESAT-6 was detected with an enzyme-linked chemiluminescent immunoassay. The recruits were also intradermally injected with PPD to assess 72-hour skin induration (the PPD skin test). Of the 892 participants, 450 (50.4%) had a positive PPD skin test and 244 (27.4%) had a positive whole-blood IFNγ release assay. Of 442 PPD-negative subjects, 88 (19.9%) had a positive whole-blood IFNγ test. Of 450 PPD-positive subjects, 156 (34.7%) had a positive whole-blood IFNγ test. Of 132 strongly PPD-positive subjects, 62 (47.0%) had a positive whole-blood IFNγ test. The agreement between the two tests was 57.2%. Of the 892 participants, 579 (64.9%) had clear vaccination scars on their arms, and of these, 382 (66.0%) had positive PPD skin responses and 162 (28.0%) were positive for the whole-blood IFNγ test. The new whole-blood IFNγ release assay might be a better indicator of LTBI than the PPD test in China.
    Molecular Diagnosis & Therapy 12/2011; 15(6):341-6. · 2.59 Impact Factor
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    ABSTRACT: China is a country with high latent tuberculosis infection (LTI) and has a policy of routine BCG vaccination. To monitor tuberculosis (TB) infection, a total of 907 new recruits to the army were interviewed and routinely examined by chest radiographs. They were intradermally injected with purified protein derivative (PPD) and detected with enzyme-linked immunospot (ELISPOT) assay with recombinant CFP-10/ESAT-6 (rCFP-10/ESAT-6) fusion protein, as a stimulus, from December 2009 to February 2010. The prevalence of LTI among new recruits was estimated as 50.2% and 30.7% by PPD skin test and ELISPOT assay, respectively. Of 452 PPD-negative and 455 PPD-positive volunteers, 132 (29.2%) and 146 (32.1%) were ELISPOT positive, respectively. The overall consistency between these two tests was 51.4% (466/907). Although 65.6% of PPD-positive volunteers and 27.8% of ELISPOT-positive volunteers (spot-forming cells; SFC 12.2 ± 21.2) could find the vaccination scars on their arms, 21.3% of PPD-positive volunteers and 35.9% of ELISPOT-positive volunteers (SFC 18.3 ± 34.6) could not. Thus, the TB infection rate in army recruits in China was not as high as previously reported. The results suggest that the ELISPOT technique, we presented, is an accurate method for screening TB infection in China.
    Apmis 06/2011; 119(6):377-84. · 2.07 Impact Factor
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    ABSTRACT: The rapid diagnosis of smear-negative pulmonary tuberculosis (TB) and extrapulmonary TB is a significant problem in clinical practice. We evaluated the usefulness of a homemade enzyme-linked immunospot (ELISPOT) assay for the diagnosis of active TB in China. Seventy-eight healthy volunteers, 60 patients with active TB, and 32 patients with non-TB diseases were evaluated by tuberculin skin test (TST), an ELISPOT assay using a recombinant CFP-10/ESAT-6 fusion protein (rCFP-10/ESAT-6) as a stimulant, and T-SPOT-TB assay. The spot-forming cells (SFC) from 78 healthy subjects containing both PPD-positive and -negative persons was 3.7 ± 6.5. Among 31 diagnosed TB patients, the ELISPOT assay had a sensitivity of 67.7%, compared to a sensitivity of 77.4% for the T-SPOT-TB assay. The ELISPOT assay was more sensitive in smear-positive TB cases (76.9%) than in smear-negative TB cases (61.1%), while T-SPOT-TB had roughly similar sensitivities in smear-positive (76.9%) and smear-negative TB cases (77.8%). The specificity was 90.6% for ELISPOT and 78.1% for T-SPOT-TB among 32 subjects with non-TB diseases. The SFC of TB cases was significantly higher than that of non-TB disease cases, and the SFC of smear-positive TB cases was significantly higher than that of smear-negative TB cases (P < 0.01). We confirmed that the homemade ELISPOT assay appears more specific for the diagnosis of active TB than T-SPOT-TB. ELISPOT assay may be a useful method for the rapid diagnosis of active TB, especially for cases of smear-negative TB.
    Molecular Biotechnology 01/2011; 47(1):18-25. · 2.26 Impact Factor
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    ABSTRACT: To study the possible relationship between polymorphic N-acetyltransferase 2 (NAT2) acetylator status and antituberculosis drug-induced hepatotoxicity and to elucidate the molecular mechanism of antituberculosis drug-induced hepatotoxicity. Blood samples from 101 tuberculosis cases with antituberculosis drug-induced hepatotoxicity and from 107 tuberculosis without antituberculosis drug-induced hepatotoxicity were collected for a case-control study. DNA of the subjects was extracted and amplified by polymerase chain reaction (PCR). The single nucleotide polymorphisms of NAT2 were determined by direct PCR sequencing. The genotype frequencies were compared between cases and controls by χ(2) test, using SPSS 12.0 software, and the association between the disease and genotypes was analyzed. Among the 101 patients with antituberculosis drug-induced liver injury, 36 patients (35.6%) were found with 282 T/T, 12 (11.9%) with 590 A/A, and 48 (47.5%) with 857 G/A or A/A. However, among the 107 controls, 9 patients (8.4%) were found with 282 T/T, 3 (2.8%) with 590 A/A, and 33 (33.8%) with 857 G/A or A/A. The patients with 282 T/T, 590 A/A, or 857 G/A or A/A genotype had a higher risk of antituberculosis drug-induced hepatotoxicity than those with 282 C/C or C/T, 590 G/G or G/A, or 857 G/G, and the OR values were 6.03 (95%CI: 2.88 - 12.62; χ(2) = 22.73, P < 0.05), 4.67 (95%CI: 1.42 - 15.44; χ(2) = 6.40, P < 0.05) and 2.03 (95%CI: 1.16 - 3.57; χ(2) = 6.08, P < 0.05) respectively. There were 40 patients with slow acetylator (39.6%) in cases with hepatotoxicity and 13 with slow acetylator (12.2%) in controls without hepatotoxicity. Patients with slow acetylator genotype (OR = 4.74, 95% CI = 2.42 - 9.28; χ(2) = 20.62, P < 0.05) had a significantly higher risk of antituberculosis drug-induced hepatotoxicity than those with rapid or intermediate acetylator genotypes. Among the cases, 19.8% (20/101) were found with NAT2(*)6A/7B, and 11.9% (12/101) with NAT2(*)6A/6A, whereas among the controls, 2.8% (3/107) were found with NAT2(*)6A/7B, and 2.8% (3/107) with NAT2(*)6A/6A respectively, the patients with NAT2(*)6A/7B and NAT2(*)6A/6A had a much higher risk of antituberculosis drug-induced hepatotoxicity, and the OR values were 8.40 (95%CI: 2.85 - 24.73; χ(2) = 14.90, P < 0.05) and 4.67 (95%CI: 1.42 - 15.44; χ(2) = 6.40, P < 0.05) respectively. Perhaps, the slow acetylation genotypes of NAT2 were the main risk factors of developing antituberculosis drug-induced hepatotoxicity.
    Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine] 01/2011; 45(1):36-40.
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    ABSTRACT: Although the delayed-type hypersensitivity (DTH) skin reaction to tuberculin is used worldwide for tuberculosis (TB) detection, it has poor diagnostic specificity due to the presence of common antigens in tuberculin shared by many mycobacterial species. The problem is noticed, especially in countries where the Bacillus Calmette-Gue´rin (BCG) vaccination is widely practiced. Thus, a new skin test antigen specific for the diagnosis of Mycobacterium tuberculosis (MTB) infection is urgently needed. CFP-10, a mycobacterial secretary protein that is absent in Mycobacterium bovis BCG and most other mycobacterial species including Mycobacterium avium, Mycobacterium intracellulare, has been shown to elicit cellular immune responses in MTB infected individuals and can be a good candidate for MTB specific diagnosis. We prepared recombinant MTB CFP-10, rCFP-10, and its utility as specific antigen for TB diagnosis was evaluated by skin testing in guinea pigs sensitized with M. tuberculosis, M. bovis, and M. bovis BCG. Our results show that the purified MTB rCFP-10 antigen elicits a positive skin response only in the guinea pigs sensitized with M. tuberculosis and M. bovis, and not in the animals sensitized with M. bovis BCG vaccine. The data presented in this study supports further testing of the use rCFP-10 as the specific antigen in the skin test for the diagnosis of MTB infection in humans.
    AFRICAN JOURNAL OF BIOTECHNOLOGY 11/2010; 9:7180-7185. · 0.57 Impact Factor
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    ABSTRACT: The detection of Mycobacteriumtuberculosis (MTB)-specific human antibodies has been an important diagnostic aid in the diagnosis of TB, especially for the bacterium-negative TB. The humoral antibody responses to different antigens of M.tuberculosis (MTB) are heterogeneous in active TB patients. Hence, detection of antibody responses to several MTB antigens may improve the sensitivity and specificity of serological diagnosis of active TB. Seventeen MTB antigens (38kD, 16kD, Ag85A, Ag85B, MPT32, MPT63, MPT64, Mtb39, MTB48, Mtb81, MTC28, Rv1009, ESAT6, CFP10, CFP10-ESAT6, katG, and LAM) were prepared by cloning, expression, and purification from E. coli, and their antigenicities were evaluated in the antibody responses of 210 active TB patients (103 sera from smear- or culture-positive patients, and 107 from smear- or culture-negative patients) and 192 healthy control (95 sera from purified protein derivative-negative healthy donors, and 97 sera from BCG-vaccinated individuals) by an enzyme-linked immunosorbent assay (ELISA). The levels of antibodies against these antigens in bacterium-negative TB patients were significantly higher than that in healthy controls (p<0.001). The sensitivity with individual antigens to detect antibody responses ranged from 55.7 to 82.9%, with the specificity from 62.0 to 92.2%. Importantly, the sensitivity with five antigens (LAM, 38kD, katG, 16kD, and MPT63 or Mtb39) to detect antibody responses reached 69.5% (146/210), with a specificity of 91.1% (17/192), and the sensitivity with another five antigens (LAM, katG, 16kD, Mtb39 and Mtb81) to detect antibody responses reached 67.1% (141/210), with a specificity of 92.7% (14/192). The combination of optimal multiple antigens to detect anti-MTB antibody responses increased the sensitivity and specificity. Therefore, detection of anti-MTB antibody responses with multiple antigens may be valuable in the clinical diagnosis of TB patients.
    Clinica chimica acta; international journal of clinical chemistry 10/2010; 411(19-20):1520-8. · 2.54 Impact Factor
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    ABSTRACT: Rifampin (RFP) is a major first-line anti-tuberculosis drug. The molecular assay currently used is the detection of rpoB gene mutations in M. tuberculosis. Recently, the Rv2629 191C allele was found to have a correlation with RFP resistance, and might become a valuable marker for the detection of RFP resistance or the Beijing genotype. We studied the association among the Rv2629 gene, rpoB gene, RFP resistance and Beijing genotype in 69 M. tuberculosis clinical isolates using DNA sequencing, conventional drug susceptibility and spoligotyping. The 191C allele was present in 92.8% (64/69) isolates. Of 29 RFP-sensitive strains, none exhibited any mutations in rpoB genes, only one strain (3.4%) was found to carry the 191A allele and 28 strains (96.6%) had the 191C alleles. Of 40 RFP-resistant strains, 30 (75%) strains had rpoB gene mutations, only 4 strains (10%) carried 191A alleles, and 36 strains (90%) exhibited 191C alleles. The 191C allele was also present in INH-sensitive, SM-sensitive or EMB-sensitive isolates. Spoligotyping analysis showed 8 distinct spoligotyping patterns. 81.1% (30/37) strains were divided into one big cluster, which had a characteristic of the Beijing genotype. Rv2629 191C allele was present in 93.3% (28/30) Beijing genotype strains, but also in non-Beijing genotype strains. These results indicate that high association is present between the ropB gene and RFP resistance. No association is present between the Rv2629 191C allele and RFP resistance, or between the Beijing genotype and RFP resistance.
    African journal of microbiology research 08/2010; 4:1575-1581. · 0.54 Impact Factor
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    ABSTRACT: The diagnosis of smear-negative and culture-negative patients with active tuberculosis (TB) is challenging. The detection of Mycobacterium tuberculosis-specific antibodies in human sera has been an important diagnostic aid. However, detection of antibody responses to a single antigen usually has a low sensitivity for diagnosis of TB. In this study, humoral immune responses against recombinant M. tuberculosis 38-kDa, MTB48, and CFP-10/ESAT-6 (culture filtrate protein 10/6-kDa early secreted antigen target of M. tuberculosis) antigens in 250 Chinese TB patients and 260 healthy subjects were evaluated by an enzyme-linked immunosorbent assay (ELISA). The levels of antibodies against those antigens in TB patients, even in bacterium-negative ones, were significantly higher than those in healthy subjects (P < 0.001). The serodiagnostic sensitivities to detect antibodies against individual antigens, i.e., recombinant M. tuberculosis 38-kDa, MTB48, and CFP-10/ESAT-6 antigens, in TB patients were 73.6%, 73.2%, and 60.4%, respectively, with specificities of 85.4%, 77.7%, and 73.8%, respectively. Importantly, the sensitivity to positively detect humoral responses to one of the antigens increased further. Our data suggest that the humoral immune responses to M. tuberculosis antigens in TB patients are heterogeneous. The 38-kDa, MTB48, and CFP-10/ESAT-6 antigens can be used as the cocktail antigens in the serodiagnosis of active TB, especially for smear- or culture-negative TB cases.
    Clinical and vaccine Immunology: CVI 03/2010; 17(3):372-5. · 2.60 Impact Factor
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    ABSTRACT: In China, latent tuberculosis infection (LTI) is frequent. To protect the health of soldiers and monitor TB infection, new recruits were routinely examined for LTI by tuberculin skin testing (TST). This is the first report on the extent of LTI in the Chinese army comparing TST and enzyme-linked immunospot (ELISPOT) assay. New recruits to the army were interviewed, routinely examined and injected intradermally with purified protein derivative (PPD) in March 2007. At the same time, 100 male soldier volunteers were detected with ELISPOT assay using recombinant CFP-10/ESAT-6 fusion protein (rCFP-10/ESAT-6) as a stimulus. The prevalence of LTI, as estimated by TST and ELISPOT assay, was 41% and 21% of new recruits, respectively. Vaccination scars on the arms could be found in 83% of volunteers with positive TST and 19% of volunteers with negative TST. Five individuals with strongly positive TST of whom 2 had negative ELISPOT were not given chemotherapy, and were observed for 20 months. None developed active TB. The prevalence of LTI in new recruits to the Chinese army is not as high as previously reported. ELISPOT technique may be the most accurate screening method for the TB infection in China, which was not interfered by BCG vaccination.
    Clinica chimica acta; international journal of clinical chemistry 06/2009; 405(1-2):110-3. · 2.54 Impact Factor
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    ABSTRACT: The problems of tuberculosis (TB) and its drug resistance are very severe in China. New therapeutic agents or regimens to treat multi-drug-resistant tuberculosis (MDR-TB) are urgently needed. In this study, the effects of Ag85A DNA or ESAT6/Ag85A chimeric DNA vaccines alone or in combination with rifampin (RFP) were studied for the treatment of mice with MDR-TB. Eighty female BALB/c mice infected with Mycobacterium tuberculosis clinical isolate HB361, which was resistant to high level of RFP, and low level of isoniazid (INH), were treated with the saline, plasmid vector pVAX1, RFP, HSP65 DNA, Ag85A DNA, Ag85A DNA combined with RFP, chimeric ESAT6/Ag85A DNA, chimeric ESAT6/Ag85A DNA combined with RFP, respectively. Different effects of DNA vaccines for the treatment of MDR-TB were demonstrated in this study. Compared with saline group, Ag85A DNA vaccine alone or Ag85A DNA in combination with rifampin group reduced the pulmonary and splenic bacterial loads by 0.58, 0.82 and 0.51, 0.69 logs, respectively. The pathological changes of lungs were also slight and the lesions were limited in comparison with that of the control mice in which the lesions were extensive and more necrotic changes were observed. Interestingly, the chimeric Ag85A/ESAT6 DNA vaccine showed the lower effect for the treatment of MDR-TB. Ag85A DNA vaccine played a main role for the treatment of TB and MDR-TB. We believe that this is the first report of the use of DNA vaccine in the treatment of MDR-TB, and that these data suggest that DNA vaccine was effective for the treatment of MDR-TB which might have the potential contribution for resolving this problem in developing countries.
    Vaccine 08/2008; 26(35):4536-40. · 3.49 Impact Factor
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    ABSTRACT: A PCR-reverse dot blot hybridization (RDBH) assay was developed for rapid identification of rifampin (RFP)-resistant genotypes in Mycobacterium tuberculosis clinical isolates. The assay used the rpoB gene as target and was used to evaluate 148 clinical isolates (97 RFP-resistant isolates and 51 RFP-susceptible isolates). At the same time, the isolates were subjected to DNA sequencing and conventional drug susceptibility test. One hundred and forty one (95.3%) and 136 (91.9%) of the 148 strains were correctly identified by DNA sequencing and RDBH assay, respectively. None of the 51 RFP-susceptible isolates examined had alterations in rpoB. The sensitivity and specificity of the DNA sequencing were 92.8% and 100%, and the positive predictive value (PPV) and negative predictive value (NPV) were 100% and 87.9%, respectively. The sensitivity and specificity of the RDBH assay were 87.6% and 100%, and the PPV and NPV were 100% and 81.0%, respectively. Codons 531 and 526 of the rpoB were found to be the most common sites of nucleotide substitutions. Mutations at codons 511, 513, 515, 516, 517, 518, and 533 were also found. There were two-codon mutations in four isolates. No deletion and insertion was found in the rpoB gene. These results indicate that the RDBH assay is a rapid, simple, and reliable method for routine identification of RFP resistance in M. tuberculosis.
    Molecular Biotechnology 08/2008; 41(1):1-7. · 2.26 Impact Factor

Publication Stats

91 Citations
35.11 Total Impact Points

Institutions

  • 2014
    • 309th Hospital of the PLA
      Peping, Beijing, China
    • The 251st Hospital of Chinese PLA
      Chzhantseyakou, Hebei, China
  • 2012
    • 307 Hospital of the Chinese People's Liberation Army
      Peping, Beijing, China
  • 2008
    • Chinese PLA General Hospital (301 Hospital)
      Peping, Beijing, China