Potential of multi-component antigens for tuberculosis diagnosis

Khurshid, Sana; Afzal, Madeeha; Khalid, Ruhi; Akhtar, Muhammad; Qazi, Mahmood Husain

doi:10.1016/j.biologicals.2017.04.004

Cited by 9 publications

(3 citation statements)

References 28 publications

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“…The combination can be either from a direct mixture as a cocktail or engineered at the molecular level to form a fusion or chimeric molecule. Previous reviews have agreed that using combination molecules as the detection elements produce superior diagnostic performance compared to their correspondence single antigen [ 23 , 24 , 25 , 26 ]. The study by Souza et al (2012) concluded that the combined ESAT6/MPB70/MPB83 antigens used in their enzyme immunoassay demonstrated superior diagnostic performance to ESAT-6 or MPB83 individually [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Amperometric Aptasensor Based on Diazonium Grafted Screen-Printed Carbon Electrode for Detection of CFP10 and MPT64 Biomarkers for Early Tuberculosis Diagnosis

et al. 2022

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Early diagnosis is highly crucial for life-saving and transmission management of tuberculosis (TB). Despite the low sensitivity and time-consuming issues, TB antigen detection still relies on conventional smear microscopy and culture techniques. To address this limitation, we report the development of the first amperometric dual aptasensor for the simultaneous detection of Mycobacterium tuberculosis secreted antigens CFP10 and MPT64 for better diagnosis and control of TB. The developed sensor was based on the aptamers–antibodies sandwich assay and detected by chronoamperometry through the electrocatalytic reaction between peroxidase-conjugated antibodies, H2O2, and hydroquinone. The CFP10 and MPT64 aptamers were immobilized via carbodiimide covalent chemistry over the disposable dual screen-printed carbon electrodes modified with a 4-carboxyphenyl diazonium salt. Under optimized conditions, the aptasensor achieved a detection limit of 1.68 ng mL−1 and 1.82 ng mL−1 for CFP10 and MPT64 antigens, respectively. The developed assay requires a small sample amount (5 µL) and can be easily performed within 2.5 h. Finally, the dual aptasensor was successfully applied to clinical sputum samples with the obtained diagnostic sensitivity (n = 24) and specificity (n = 13) of 100%, respectively, suggesting the readiness of the developed assay to be used for TB clinical application.

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Section: Introductionmentioning

confidence: 99%

Simultaneous Amperometric Aptasensor Based on Diazonium Grafted Screen-Printed Carbon Electrode for Detection of CFP10 and MPT64 Biomarkers for Early Tuberculosis Diagnosis

et al. 2022

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“…However, the sensitivities and specificities of the currently available options based on single or multiple target antigens for TB are variable and do not yet meet the requirements for clinical use. In 2011, the World Health Organization (WHO) issued a policy recommendation against the use of the various commercial serological tests for TB diagnosis due to the suboptimal sensitivity and specificity ( 9 ). However, further research and development in this field, specifically the identification and screening of novel serodiagnostic antigens, is still highly recommended by WHO ( 10 ).…”

Section: Introductionmentioning

confidence: 99%

Development and Evaluation of a Fusion Polyprotein Based on HspX and Other Antigen Sequences for the Serodiagnosis of Tuberculosis

Zhou

Cui

et al. 2021

Front. Immunol.

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BackgroundThe lack of suitable diagnostic tools contributes to the high prevalence of tuberculosis (TB) worldwide. Serological tests, based on multiple target antigens, represent an attractive option for diagnosis of this disease due to their rapidity, convenience, and low cost.MethodsMeasures to reduce non-specific reactions and thereby improve the specificity of serological tests were investigated, including blocking antibodies against common bacteria in serum samples and synthesizing polypeptides covering non-conserved dominant B-cell epitopes of antigens. In addition, a fusion polyprotein containing HspX and eight other antigen sequences was constructed and expressed to increase overall sensitivity of the tests.ResultsInclusion of Escherichia coli lysate partially increased the specificity of the serological tests, while synthesis and inclusion of peptides containing non-conserved sequences of TB antigens as well as dominant B-cell epitopes reduced non-specific reactions without a decrease in sensitivity of the tests. A polyprotein fusing HspX and eight other antigen sequences was constructed and displayed 60.2% sensitivity, which was higher than that of HspX and the other individual antigen segments. Moreover, the specificity of the polyprotein was 93.8%, which was not significantly decreased when compared with HspX and the other individual antigen segments.ConclusionsThe roles of the fusion polyprotein in the humoral immune response against TB infection were demonstrated and provide a potential novel approach for the development of TB diagnostics.

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“…In 2011, WHO issued a policy recommendation against the use of the various commercial serological tests for TB diagnosis due to the suboptimal sensitivity and specificity. 6 However, further research and development in this field, specifically the identification and screening of novel serodiagnostic antigens, is still highly recommended by WHO. 7 One major challenge for TB serodiagnosis was false positive reaction with healthy individuals, which reduced the specificity.…”

Section: Introductionmentioning

confidence: 99%

Assessment of a Fusion Polyprotein Based on HspX and Other Eight Antigen Segments for Tuberculosis Serodiagnosis

Zhou

Cui

et al. 2019

Preprint

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Background: The lack of suitable diagnostic tools has paved the cornerstone for controlling and treatment of tuberculosis. Serological tests, based on multiple target antigens, represent an attractive option for its rapidity, convenience, and low-cost. Methods: Measures, including blocking antibodies against common bacteria in serum samples and synthesizing polypeptides covering un-conserved dominant B-cell epitopes of antigens, were attempted to reduce nonspecific reactions and thus improved the specificity. Meanwhile, a fusion polyprotein containing HspX and other eight antigen segments was constructed and expressed to increase the whole sensitivity. Results: The addition of only E. coli lysate partly increased the specificities. Another, peptides containing un-conserved fragments of TB antigens as well as dominant B-cell epitopes were synthesized and used to effectively reduce non-specific reactions without dramatically decreasing their sensitivities. Meanwhile, a polyprotein fusing HspX and other eight antigen segments was constructed and the sensitivity of the polyprotein was 60.2%, which was higher that of HspX and other individual antigen segments. Importantly, the specificity of the polyprotein was 93.6%, which was not significantly decreased. Conclusions: Our results demonstrate the roles of fusion polyproteins in the humoral immunity against TB infection and provide a potential novel approach for TB diagnostic development.

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Potential of multi-component antigens for tuberculosis diagnosis

Cited by 9 publications

References 28 publications

Simultaneous Amperometric Aptasensor Based on Diazonium Grafted Screen-Printed Carbon Electrode for Detection of CFP10 and MPT64 Biomarkers for Early Tuberculosis Diagnosis

Simultaneous Amperometric Aptasensor Based on Diazonium Grafted Screen-Printed Carbon Electrode for Detection of CFP10 and MPT64 Biomarkers for Early Tuberculosis Diagnosis

Development and Evaluation of a Fusion Polyprotein Based on HspX and Other Antigen Sequences for the Serodiagnosis of Tuberculosis

Assessment of a Fusion Polyprotein Based on HspX and Other Eight Antigen Segments for Tuberculosis Serodiagnosis

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