Novel monoclonal antibodies to ESAT-6 and CFP-10 antigens for ELISA-based diagnosis of pleural tuberculosis

Tt, Feng; Cm, Shou; Shen, Liuhong; Qian, Yajuan; Zg, Wu; Fan, Jun; Zhang, Yanzhong; Tang, Yutao; Np, Wu; Hz, Lu; Hp, Yao

doi:10.5588/ijtld.10.0393

Cited by 25 publications

(32 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Additionally, the sensitivities of these peptides, especially in combination, were higher compared to the classical smear assay (sensitivity, 64%). Recently, monoclonal antibodies were generated against the M. tuberculosis-specific antigen ESAT-6, resulting in an ESAT-6-specific ELISA that yielded a specificity of 100% and a sensitivity of 95.4% (Feng et al 2011). These data correlate well with our findings when using this antigen.…”

Section: Discussionsupporting

confidence: 80%

Immunologic evaluation and validation of methods using synthetic peptides derived from Mycobacterium tuberculosis for the diagnosis of tuberculosis infection

Araujo¹,

Giampietro²,

Bochichio³

et al. 2013

Mem. Inst. Oswaldo Cruz

View full text Add to dashboard Cite

The World Health Organization (WHO) lists tuberculosis (TB) as the most important fatal infection worldwide (WHO 2009). The development of a new diagnostic test for TB infection is an important component of the Global Plan to Stop TB and the WHO Stop TB Strategy. In 2005 alone, an estimated 8.8 million individuals were infected with TB and 1.6 million people died of the disease (WHO 2009). Importantly, less than one half of the total 8.8 million estimated cases were diagnosed as smear-positive; the diagnosis of smear-negative patients has proven to be more challenging. Currently, a definitive diagnosis of both pulmonary TB (PTB) and extrapulmonary TB (EPTB) relies on the time consuming culture of mycobacteria. A vast majority of TB cases occur in developing countries that have limited resources. Rapid, inexpensive diagnostic tests would aid these countries in limiting the spread of infection within their communities (WHO 2009). In addition, molecular methods based on nucleic acid amplification to diagnose TB infection are rapid, highly specific and more sensitive than microscopic examination of smears, but are less sensitive compared to culture assays for smear-negative TB cases (Abebe et al. 2007).Serological tests that rely on the detection of antibodies against Mycobacterium tuberculosis-specific antigens possess several advantages: they are simple, inexpensive and feasible for the diagnosis of TB. Potential M. tuberculosis antigens were recently reviewed in a metaanalysis. A total of 254 studies were identified that encompassed nine native proteins, 27 recombinant proteins, 15 lipid-derived antigens and 30 combination antigen targets. These results indicated that highly specific tests frequently exhibited poor sensitivity, which limited the use of these antigens when a single antigen was used in the assay (Steingart et al. 2009). Recently, there has been renewed interest in the development of antibody-based diagnostic assays that utilise multiple antigens to achieve high sensitivity and specificity (Abebe et al. 2007). Many attempts to develop a serologic TB test have been made. These assays need to discriminate active from latent infection, avoid cross-reactivity with Bacillus Calmette-

show abstract

Section: Discussionsupporting

confidence: 80%

Immunologic evaluation and validation of methods using synthetic peptides derived from Mycobacterium tuberculosis for the diagnosis of tuberculosis infection

Araujo¹,

Giampietro²,

Bochichio³

et al. 2013

Mem. Inst. Oswaldo Cruz

View full text Add to dashboard Cite

show abstract

“…Serum CFP-10 and ESAT-6 expression theoretically can be used to diagnose all active Mtb infections, including EPTB cases (20); however, some NTM strains express homologs that may reduce the utility of these proteins as biomarkers (21). Since peptide sequence is the gold standard for protein discrimination, we examined whether tryptic peptides could distinguish Mtbderived ESAT-6 and CFP-10 from homologs produced by other species.…”

Section: Resultsmentioning

confidence: 99%

Quantification of circulating Mycobacterium tuberculosis antigen peptides allows rapid diagnosis of active disease and treatment monitoring

Liu

Zhao

Fan

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Tuberculosis (TB) is a major global health threat, resulting in an urgent unmet need for a rapid, non-sputum-based quantitative test to detect active Mycobacterium tuberculosis (Mtb) infections in clinically diverse populations and quickly assess Mtb treatment responses for emerging drug-resistant strains. We have identified Mtb-specific peptide fragments and developed a method to rapidly quantify their serum concentrations, using antibody-labeled and energy-focusing porous discoidal silicon nanoparticles (nanodisks) and high-throughput mass spectrometry (MS) to enhance sensitivity and specificity. NanoDisk-MS diagnosed active Mtb cases with high sensitivity and specificity in a case-control study with cohorts reflecting the complexity of clinical practice. Similar robust sensitivities were obtained for cases of culture-positive pulmonary TB (PTB; 91.3%) and extrapulmonary TB (EPTB; 92.3%), and the sensitivities obtained for culture-negative PTB (82.4%) and EPTB (75.0%) in HIV-positive patients significantly outperformed those reported for other available assays. NanoDisk-MS also exhibited high specificity (87.1-100%) in both healthy and high-risk groups. Absolute quantification of serum Mtb antigen concentration was informative in assessing responses to antimycobacterial treatment. Thus, a NanoDisk-MS assay approach could significantly improve the diagnosis and management of active TB cases, and perhaps other infectious diseases as well.D espite international efforts and initiatives, tuberculosis (TB) remains a major public health concern worldwide, associated with high morbidity and mortality (1, 2). Detecting active TB cases and monitoring their responses to therapy are fraught with challenges, relying predominantly on microbiologic techniques that use sputum samples, including acid-fast Bacillus (AFB) smear microscopy-widely used as an initial test for TB diagnosis (3, 4)-and Mycobacterium tuberculosis (Mtb) culture, both of which have only moderate sensitivity and specificity and a long turnaround time (5, 6). Moreover, sputum samples are difficult to obtain after symptom improvement, and often are not diagnostically useful for extrapulmonary TB (EPTB) cases. The PCR-based Xpert MTB/RIF sputum assay was introduced to improve the speed and specificity of TB diagnosis, but this assay has poor sensitivity under low bacterial loads and cannot distinguish live and nonviable Mtb contributions (7,8). A recent World Health Organization (WHO) policy update acknowledged the low quality of evidence supporting the use of Xpert MTB/RIF to diagnose EPTB (9). Diagnostic challenges can be further magnified in patients coinfected with HIV and TB (10). In addition, none of these techniques provides quantitative results that can be used to monitor treatment effects (11,12).Consequently, there is an urgent need, highlighted as a high priority in a recent WHO consensus report (13), for the development of rapid, quantitative, non-sputum-based biomarker tests that do not require bacterial isolation to detect active TB (13). Non-s...

show abstract

“…The CFP-10 protein, encoded by the Rv3874 ( esxB ) gene, and the ESAT-6 protein, encoded by the Rv3875 ( esxA ) gene, can interact in vitro to form a tight 1∶1 heterodimer [16], [17]. These two proteins are potent T-cell antigens recognised by over 70% of tuberculosis patients and are thus good TB biomarkers [18], [19], [20]. Moreover, CFP-10 and ESAT-6 are not present in many nontuberculous mycobacteria and in the Mycobacterium bovis Bacillus Calmette-Guérin (BCG) vaccine [5], [18], [19].…”

Section: Introductionmentioning

confidence: 99%

Selection and Application of ssDNA Aptamers to Detect Active TB from Sputum Samples

et al. 2012

View full text Add to dashboard Cite

BackgroundDespite the enormous global burden of tuberculosis (TB), conventional approaches to diagnosis continue to rely on tests that have major drawbacks. The improvement of TB diagnostics relies, not only on good biomarkers, but also upon accurate detection methodologies. The 10-kDa culture filtrate protein (CFP-10) and the 6-kDa early secreted antigen target (ESAT-6) are potent T-cell antigens that are recognised by over 70% of TB patients. Aptamers, a novel sensitive and specific class of detection molecules, has hitherto, not been raised to these relatively TB-specific antigens.MethodsDNA aptamers that bind to the CFP-10.ESAT-6 heterodimer were isolated. To assess their affinity and specificity to the heterodimer, aptamers were screened using an enzyme-linked oligonucleotide assay (ELONA). One suitable aptamer was evaluated by ELONA using sputum samples obtained from 20 TB patients and 48 control patients (those with latent TB infection, symptomatic non TB patients, and healthy laboratory volunteers). Culture positivity for Mycobacterium tuberculosis (Mtb) served as the reference standard. Accuracy and cut-points were evaluated using ROC curve analysis.ResultsTwenty-four out of the 66 aptamers that were isolated bound significantly (p<0.05) to the CFP-10.ESAT-6 heterodimer and six were further evaluated. Their dissociation constant (KD) values were in the nanomolar range. One aptamer, designated CSIR 2.11, was evaluated using sputum samples. CSIR 2.11 had sensitivity and specificity of 100% and 68.75% using Youden’s index and 35% and 95%, respectively, using a rule-in cut-point.ConclusionThis preliminary proof-of-concept study suggests that a diagnosis of active TB using anti-CFP-10.ESAT-6 aptamers applied to human sputum samples is feasible.

show abstract

Novel monoclonal antibodies to ESAT-6 and CFP-10 antigens for ELISA-based diagnosis of pleural tuberculosis

Abstract: The ESAT-6 and CFP-10 ELISAs incorporating mAbs generated in this study serve as potential tools in the laboratory diagnosis of TB.

Cited by 25 publications

References 0 publications

Immunologic evaluation and validation of methods using synthetic peptides derived from Mycobacterium tuberculosis for the diagnosis of tuberculosis infection

Immunologic evaluation and validation of methods using synthetic peptides derived from Mycobacterium tuberculosis for the diagnosis of tuberculosis infection

Quantification of circulating Mycobacterium tuberculosis antigen peptides allows rapid diagnosis of active disease and treatment monitoring

Selection and Application of ssDNA Aptamers to Detect Active TB from Sputum Samples

Contact Info

Product

Resources

About