Detection of <i>Mycobacterium tuberculosis</i> lipoarabinomannan and CFP-10 (Rv3874) from urinary extracellular vesicles of tuberculosis patients by immuno-PCR

Dahiya, Bhawna; Khan, Anish; Mor, Preeti; Kamra, Ekta; Singh, Netrapal; Gupta, K. B.; Sheoran, Anita; Sreenivas, Vishnubhatla; Mehta, Promod K

doi:10.1093/femspd/ftz049

Cited by 37 publications

(42 citation statements)

References 49 publications

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“…The rate of progression and the mortality rate for children and adults with extrapulmonary disseminated disease are higher than patients with isolated pulmonary TB 10 . These populations cannot produce sputum for conventional diagnostic modalities, thus an accurate urine screening test for active TB in these patients is critically needed 2 – 4 , 10 – 12 . Previous studies of urinary LAM diagnostics conducted on pediatric TB patients yielded low sensitivity, but nevertheless showed the promise of extending urinary LAM testing from adults to children 13 , 14 .…”

Section: Introductionmentioning

confidence: 99%

Lipoarabinomannan antigenic epitope differences in tuberculosis disease subtypes

Magni

Rruga

Alsaab

et al. 2020

Sci Rep

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An accurate urine test for diverse populations with active tuberculosis could be transformative for preventing TB deaths. Urinary liporabinomannan (LAM) testing has been previously restricted to HIV co-infected TB patients. In this study we evaluate urinary LAM in HIV negative, pediatric and adult, pulmonary and extrapulmonary tuberculosis patients. We measured 430 microbiologically confirmed pretreatment tuberculosis patients and controls from Peru, Guinea Bissau, Venezuela, Uganda and the United States using three monoclonal antibodies, MoAb1, CS35, and A194, which recognize distinct LAM epitopes, a one-sided immunoassay, and blinded cohorts. We evaluated sources of assay variability and comorbidities (HIV and diabetes). All antibodies successfully discriminated TB positive from TB negative patients. ROAUC from the average of three antibodies' responses was 0.90; 95% CI 0.87-0.93, 90% sensitivity, 73.5% specificity (80 pg/mL). MoAb1, recognizing the 5-methylthio-dxylofuranose(MTX)-mannose(Man) cap epitope, performed the best, was less influenced by glycosuria and identified culture positive pediatric (N = 19) and extrapulmonary (N = 24) patients with high accuracy (ROAUC 0.

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Section: Introductionmentioning

confidence: 99%

Lipoarabinomannan antigenic epitope differences in tuberculosis disease subtypes

Magni

Rruga

Alsaab

et al. 2020

Sci Rep

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“…Moreover, urine is quite easy to obtain that has a lesser risk of infection to the laboratory workers but it carries low concentration of M. tuberculosis biomarkers [18]. Detection of mycobacterial lipoarabinomannan (LAM) within urine samples is considered as an attractive biomarker, which has been demonstrated in HIV-coinfected and HIVuninfected TB individuals by lateral flow immunochromatography and ELISA methods though poor sensitivities were observed in HIV-uninfected TB individuals [17,19]. However, urinary extracellular vesicles (EVs) isolated from TB patients can furnish intact pathogen-derived biomarkers and also concentrate those biomarkers [19].…”

Section: Immuno-pcrmentioning

confidence: 99%

“…Detection of mycobacterial lipoarabinomannan (LAM) within urine samples is considered as an attractive biomarker, which has been demonstrated in HIV-coinfected and HIVuninfected TB individuals by lateral flow immunochromatography and ELISA methods though poor sensitivities were observed in HIV-uninfected TB individuals [17,19]. However, urinary extracellular vesicles (EVs) isolated from TB patients can furnish intact pathogen-derived biomarkers and also concentrate those biomarkers [19]. We recently detected CFP-10 and LAM from the urinary EVs of PTB and EPTB patients by I-PCR, which revealed significantly higher sensitivities than ELISA [19].…”

Section: Immuno-pcrmentioning

confidence: 99%

“…However, urinary extracellular vesicles (EVs) isolated from TB patients can furnish intact pathogen-derived biomarkers and also concentrate those biomarkers [19]. We recently detected CFP-10 and LAM from the urinary EVs of PTB and EPTB patients by I-PCR, which revealed significantly higher sensitivities than ELISA [19]. Furthermore, sensitivities attained with LAM detection by I-PCR were better than CFP-10 detection, but urinary EVs of UGTB patients were not examined.…”

Section: Immuno-pcrmentioning

confidence: 99%

“…Furthermore, sensitivities attained with LAM detection by I-PCR were better than CFP-10 detection, but urinary EVs of UGTB patients were not examined. To further improve the diagnosis of UGTB, we recommend detection of a cocktail of M. tuberculosis biomarkers, for example, LAM+CFP-10/MPT64 within urinary EVs of UGTB patients by I-PCR/nanoparticle-based I-PCR assay, which may match the WHO criteria of high-priority target product profile [19].…”

Section: Immuno-pcrmentioning

confidence: 99%

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Recent Trends in Diagnosis of Urogenital Tuberculosis

Mehta

Kamra

2020

Future Microbiol.

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the efficient diagnosis of UGTB by reliable tests would certainly reduce the infertility in both men and women and the other sequelae associated with disease. " Urogenital tuberculosis (UGTB) is the second most common form of extrapulmonary TB (EPTB) in developing nations with severe epidemic situations. For example, India (comprising 45% of EPTB cases) and the third most common form in locations with a lower incidence of TB [1,2], which include both renal and genital TB. However, concomitant UGTB and pulmonary TB (PTB) are found in 2-10 and 15-20% of patients in developed and developing nations, respectively [2]. In fact, UGTB occurs after a prolonged period of latent infection followed by hematogenous spread to the kidneys, fallopian tubes and epididymis [3]. The clinical manifestations of female UGTB include primary or secondary infertility, menstrual dysfunction and pelvic inflammatory disease [4]. Infertility is the most common complication, which occurs in 5-16% of Indian women caused by irreversible damage to the fallopian tubes. Mostly, fallopian tubes are associated with congestion and miliary tubercles, while endometrium is linked with caseation and ulceration in 50-80% cases, resulting in Asherman's syndrome [5]. Similarly, the most frequent sites for male UGTB are the epididymis and prostate followed by seminal vesicles and testicles originating from renal foci, which also lead to infertility [6]. Owing to asymptomatic nature and varied clinical presentations (ranging from vague urinary symptoms to chronic kidney disease) that mimic several urologic and genital diseases, diagnosis of UGTB is a daunting challenge [3].Smear/culture, histopathology, interferon-γ release assays (IGRAs), intravenous pyelography, laparoscopy and nucleic acid amplification tests (NAATs) are the main modalities employed in the diagnosis of UGTB [6][7][8], which have certain limitations. Smear microscopy and culture for Mycobacterium tuberculosis identification (on Lowenstein-Jensen medium or BACTEC-460/MGIT-960) are commonly used methods but they lead to poor sensitivities due to paucibacillary nature of specimens [7]. Though urine culture is considered the gold standard, it requires skillful technicians and has a prolonged turnaround time of approximately 6-8 weeks [7] when irreparable damage to the fallopian tubes and epididymis already occurs. Delayed diagnosis also leads to nonfunctioning unilateral kidney, contracted bladder and renal failure [1]. Endoscopic procedures such as laparoscopy/hysteroscopy are often linked with operative risks and mostly cause flaring of infection [5], while histopathological examination can be confirmatory only when it demonstrates acid-fast bacilli in the tissues that are very scarce in endometrial biopsies (EBs) [7]. Intravenous pyelography may be suggestive of UGTB but it is nonspecific and often mimics other pathologic lesions [1]. IGRAsImmunological procedures such as IGRAs are widely employed for the early detection of UGTB cases. Notably, Kim et al. [8] reported a sensitivity of 52.6%...

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Monocrystalline Labeling Enables Stable Plasmonic Enhancement for Isolation‐Free Extracellular Vesicle Analysis

Wang

Zheng

Wang

et al. 2022

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such applications due to their straightforward synthesis, optical properties, and the diverse array of surface chemistry modifications that can be used to functionalize them with factors that recognize specific biomolecules and thereby generate probes that can detect and quantify specific targets of interest. [2] Such NP probes are typically highly stable, the signal they produce is durable, and can be more intense than that produced by fluorescent dyes or quantum dots, yet can be read by inexpensive dark field microscopes available in most research and clinical laboratories. [3] Scattering signal corresponds to NP size, although the inverse relationship between NP signal and NP binding kinetics, can constrain the size range of NPs used in nanoprobe applications and their limits of detection. Alternate detection approaches that employ enzymatic reactions to enhance signal intensity (e.g., an electrochemical-chemical-enzymatic redox cycling reaction read by an electrochemical sensor [4] and conversion of enzyme substrate to an insoluble form read by a simple surface plasmonic resonance sensor [5] ) can be more sensitive but may need to be read in real-time, require sophisticated equipment and highly trained personnel, or depend upon close temperature control during transport, storage, and assay performance.The signal produced by NP probes are a function of their composition, geometry, and size but can be modified in situ by increasing the apparent size or surface composition of affinity bound NPs through specific NP growth, modification, or aggregation methods. Multiple methods have been developed to enhance biosensing characteristics of localized surface plasmonic resonance (LSPR) probes, but most of these involve tradeoffs between signal enhancement efficiency, target specificity, and assay complexity. For example, NPs with complex (e.g., nanostars) or hybrid compositions (e.g., gold/silica structures) can exhibit enhanced light scattering characteristics, but these material can be difficult to synthesis and manipulate, [6] and increasing NP size to improve scattering can negatively affect their binding kinetics. [7] However, chemical reactions employed to induce in situ NP growth or surface modifications after their binding to a biomarker target can disrupt probetarget interactions to produce signal loss or allow nonspecific deposition of NP material on other matrixes present the Sensitive detection of extracellular vesicles (EVs) as emerging biomarkers has shown great promises for disease diagnosis. Plasmonic metal nanostructures conjugated with molecules that bind specific biomarker targets are widely used for EVs sensing but involve tradeoffs between particle-sizedependent signal intensity and conjugation efficiency. One solution to this problem would be to induce nucleation on nanoparticles that have successfully bound a target biomarker to permit in situ nanoparticle growth for signal amplification, but approaches that are evaluated to date require harsh conditions or lack nucleation specificity, prohibi...

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Detection of Mycobacterium tuberculosis lipoarabinomannan and CFP-10 (Rv3874) from urinary extracellular vesicles of tuberculosis patients by immuno-PCR

Cited by 37 publications

References 49 publications

Lipoarabinomannan antigenic epitope differences in tuberculosis disease subtypes

Lipoarabinomannan antigenic epitope differences in tuberculosis disease subtypes

Recent Trends in Diagnosis of Urogenital Tuberculosis

Monocrystalline Labeling Enables Stable Plasmonic Enhancement for Isolation‐Free Extracellular Vesicle Analysis

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