Preparation of immunochromatographic strips for rapid detection of early secreted protein ESAT-6 and culture filtrate protein CFP-10 from Mycobacterium tuberculosis

Wu, Xiaoxin; Wang, Yeping; Weng, Tian-Hao; Hu, Chen-Yu; Wang, Frederick X.C.; Wu, Zhigang; Yu, Dongshan; Lu, Huoquan; Yao, Hang‐Ping

doi:10.1097/md.0000000000009350

Cited by 11 publications

(12 citation statements)

References 32 publications

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“…ICS have also been developed for the detection of some bacterial pathogens, in particular foodborne pathogens and their toxins [ 46 ]. Development of ICS for the detection of human pathogens such as Mycobacterium tuberculosis [ 47 ], S. aureus [ 26 , 48 ], Helicobacter pylori [ 49 ], H. influenzae [ 50 ], and Neisseria gonorrhoeae [ 51 ] has been attempted, yet not as extensively as the previous applications.…”

Section: Discussionmentioning

confidence: 99%

Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of Klebsiella pneumoniae Causing Neonatal Sepsis

et al. 2021

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Neonatal sepsis is a leading cause of death among newborns and infants, especially in the developing world. The problem is compounded by the delays in pinpointing the causative agent of the infection. This is reflected in increasing mortality associated with these cases and the spread of multi-drug-resistant bacteria. In this work, we deployed bioinformatics and proteomics analyses to determine a promising target that could be used for the identification of a major neonatal sepsis causative agent, Klebsiella pneumoniae. A 19 amino acid peptide from a hypothetical outer membrane was found to be very specific to the species, well conserved among its strains, surface exposed, and expressed in conditions simulating infection. Antibodies against the selected peptide were conjugated to gold nanoparticles and incorporated into an immunochromatographic strip. The developed strip was able to detect as low as 105 CFU/mL of K. pneumoniae. Regarding specificity, it showed negative results with both Escherichia coli and Enterobacter cloacae. More importantly, in a pilot study using neonatal sepsis cases blood specimens, the developed strip selectively gave positive results within 20 min with those infected with K. pneumoniae without prior sample processing. However, it gave negative results in cases infected with other bacterial species.

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Section: Discussionmentioning

confidence: 99%

Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of Klebsiella pneumoniae Causing Neonatal Sepsis

et al. 2021

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“…(11,12). Previous studies have employed ELISA to detect CFP-10 and ESAT-6 (13,14) or MPT-64 (15)(16)(17) for TB diagnosis, primarily in non-serum samples, but most of these reports lack follow-up studies to validate their results or to evaluate the utility of these assays for EPTB diagnosis. SCE assay analysis revealed 85.5% overall sensitivity for Confirmed TB and Unconfirmed TB cases and 100% specificity to exclude individuals with non-TB diagnoses.…”

Section: Diagnostic Performance Of Different Diagnostic Modelsmentioning

confidence: 99%

Serum-Based Diagnosis of Pediatric Tuberculosis by Assay of Mycobacterium tuberculosis Factors: a Retrospective Cohort Study

Lyon

Nguyen

et al. 2021

J Clin Microbiol

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Diagnosis of pediatric tuberculosis (TB) is often complicated by its non-specific symptoms, paucibacillary nature, and the need for invasive specimen collection techniques. However, a recently reported assay that detects Mycobacterium tuberculosis (Mtb) virulence factors in serum can diagnose various TB manifestations, including paucibacillary TB cases, in adults with good sensitivity and specificity. The current study examined the ability of this Mtb biomarker assay to diagnose pediatric TB using archived cryopreserved serum samples drawn from children ≤ 18 years who were screened for suspected TB as part of a prospective population-based active surveillance study. In this analysis, any detectable level of the Mtb virulence factors CFP-10 or ESAT-6 was considered direct evidence of TB. Serum samples were analyzed from 105 children evaluated for TB (55 TB cases and 50 close contacts without TB). Results of this analysis yielded sensitivity of 85·5% (95% CI 73·3–93·5). Similar diagnostic sensitivity was observed for culture-positive (87·5; 95% CI: 67·6–97·3) and culture-negative (83·9; 95% CI: 66·3–94·5) TB cases, and for culture negative pulmonary (77·8%; 95% CI: 40·0–97·2) and extrapulmonary (86·4%; 95% CI: 65·1–97·1) TB cases. These results suggest that serum biomarker analysis holds significant promise for rapid and sensitive diagnosis of pediatric TB cases, including extrapulmonary or paucibacillary TB cases. The ability to use frozen samples for this analysis should also permit assays to be performed at central sites, without a requirement for strict timelines for sample analysis.

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“…This test, however, was found to be more suitable for TB testing in HIV-positive patients. The commercially available LFIA-based TB RDTs have limited endorsement from the World Health Organization (WHO) [ 10 ], and in some studies, the reported sensitivity and specificity of the diagnostic kits were inconsistent [ 11 ]. The poor specificity may be attributed to cross-reactivity with the BCG vaccine and other Mycobacterium species [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Development of Gold-Nanoparticle-Based Lateral Flow Immunoassays for Rapid Detection of TB ESAT-6 and CFP-10

et al. 2023

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The current study reports on the development of a rapid and cost-effective TB-antigen diagnostic test for the detection of Mycobacterium biomarkers from non-sputum-based samples. Two gold nanoparticle (AuNP)-based rapid diagnostic tests (RDTs) in the form of lateral flow immunoassays (LFIAs) were developed for detection of immunodominant TB antigens, the 6 kDa early secreted antigen target EsxA (ESAT-6) and the 10 kDa culture filtrate protein EsxB (CFP-10). AuNPs were synthesized using the Turkevich method and characterized by UV-vis spectrophotometer and transmission electron microscope (TEM). The AuNP–detection probe conjugation conditions were determined by comparing the stability of 14 nm AuNPs at different pH conditions, following salt challenge. Thereafter, ESAT-6 and CFP-10 antibodies were conjugated to the AuNPs and used for the colorimetric detection of TB antigens. Selection of the best detection and capture antibody pairs was determined by Dot spotting. The limits of detection (LODs) for the LFIAs were evaluated by dry testing. TEM results showed that the 14 nm AuNPs were mostly spherical and well dispersed. The ESAT-6 LFIA prototype had an LOD of 0.0625 ng/mL versus the CFP-10 with an LOD of 7.69 ng/mL. Compared to other studies in the literature, the LOD was either similar or lower, outperforming them. Moreover, in some of the previous studies, an enrichment/extraction step was required to improve on the LOD. In this study, the LFIAs produced results within 15 min and could be suitable for use at PoCs either in clinics, mobile clinics, hospitals or at home by the end user. However, further studies need to be conducted to validate their use in clinical samples.

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Preparation of immunochromatographic strips for rapid detection of early secreted protein ESAT-6 and culture filtrate protein CFP-10 from Mycobacterium tuberculosis

Abstract: Supplemental Digital Content is available in the text

Cited by 11 publications

References 32 publications

Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of Klebsiella pneumoniae Causing Neonatal Sepsis

Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of Klebsiella pneumoniae Causing Neonatal Sepsis

Serum-Based Diagnosis of Pediatric Tuberculosis by Assay of Mycobacterium tuberculosis Factors: a Retrospective Cohort Study

Development of Gold-Nanoparticle-Based Lateral Flow Immunoassays for Rapid Detection of TB ESAT-6 and CFP-10

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