A Homogeneous Immunoassay Method for Detecting Interferon-Gamma in Patients with Latent Tuberculosis Infection
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IFN-γ release assays (IGRAs) have been developed as viable alternative diagnostic tools for detecting latent tuberculosis infection (LTBI). A customized homogeneous sandwich luminescent oxygen channeling immunoassay (LOCI) was used to quantify IFN-γ levels in IGRAs. Samples were collected from healthy volunteers (n = 40) who were T-Spot-negative and T-Spot-positive patients (n = 32) at rest. Then the amount of IFN-γ in the supernatant of IGRAs was measured by LOCI. The results demonstrated a low background, and high sensitivity, specificity, accuracy, and reproducibility, and a short assay time (only 30 min) with LOCI for IFN-γ. The recovery range was 81.63-102.06%, the coefficients of variation were below 5%, and the limit of detection was 19.0 mIU/ml. Excellent agreement between LOCI IFN-γ and the T-SPOT.TB test was obtained (97.2% agreement, κ = 0.94). The LOCI IFN-γ concentrations were significantly higher in T-Spot-positive patients than in the healthy group (p < 0.001). Moreover, as observed for the comparative LOCI IFN-γ assay, IFN-γ concentrations were related to the numbers of T-SPOT.TB spots. We have established an in vitro blood test for LTBI diagnosis, defined as LOCI IFN-γ. A high level of agreement between the LOCI IFN-γ method and T-SPOT.TB assay was observed in clinical studies that showed the LOCI IFN-γ method could determine LTBI. This study shows acceptable performance characteristics of the LOCI IFN-γ assay to diagnose LTBI.
Abstract
IFN-γ release assays (IGRAs) have been developed as viable alternative diagnostic tools for detecting latent tuberculosis infection (LTBI). A customized homogeneous sandwich luminescent oxygen channeling immunoassay (LOCI) was used to quantify IFN-γ levels in IGRAs. Samples were collected from healthy volunteers (n = 40) who were T-Spot-negative and T-Spot-positive patients (n = 32) at rest. Then the amount of IFN-γ in the supernatant of IGRAs was measured by LOCI. The results demonstrated a low background, and high sensitivity, specificity, accuracy, and reproducibility, and a short assay time (only 30 min) with LOCI for IFN-γ. The recovery range was 81.63-102.06%, the coefficients of variation were below 5%, and the limit of detection was 19.0 mIU/ml. Excellent agreement between LOCI IFN-γ and the T-SPOT.TB test was obtained (97.2% agreement, κ = 0.94). The LOCI IFN-γ concentrations were significantly higher in T-Spot-positive patients than in the healthy group (p < 0.001). Moreover, as observed for the comparative LOCI IFN-γ assay, IFN-γ concentrations were related to the numbers of T-SPOT.TB spots. We have established an in vitro blood test for LTBI diagnosis, defined as LOCI IFN-γ. A high level of agreement between the LOCI IFN-γ method and T-SPOT.TB assay was observed in clinical studies that showed the LOCI IFN-γ method could determine LTBI. This study shows acceptable performance characteristics of the LOCI IFN-γ assay to diagnose LTBI.
참고문헌 (23)
- Barry CE, "The spectrum of latent tuberculosis: rethinking the biology and intervention strategies", Nat. Rev. Microbiol, 7, pp.845~855 (2009)
- Beaudet L, "Homogeneous assays for single-nucleotide polymorphism typing using AlphaScreen", Gen. Res, 11, pp.600~608 (2001)
- Bosse R, "Miniaturizing screening: how low can we go today", Drug Discov. Today, pp.42~47 (2000)
- Cohn DL, "Targeted tuberculin testing and treatment of latent tuberculosis infection", MMWR Morb. Mortal. Wkly. Rep, 49, pp.1~54 (2000)
- Connell TG, "Performance of a whole blood interferon gamma assay for detecting latent infection with Mycobacterium tuberculosis in children", Thorax, 61, pp.616~620 (2006)
- Diel R, "Predictive value of interferon-gamma release assays and tuberculin skin testing for progression from latent TB infection to disease state: a meta-analysis", Chest, 142, pp.63~75 (2012)
- Eglen RM, "The use of AlphaScreen technology in HTS: current status", Curr. Chem. Gen, 1, pp.2~10 (2008)
- Fietta A, "Comparison of a whole-blood interferon-γ assay and tuberculin skin testing in patients with active tuberculosis and individuals at high or low risk of Mycobacterium tuberculosis infection", Am. J. Infect. Control, 31, pp.347~353 (2003)
- Goletti D, "Interferon-gamma release assays for the diagnosis of latent Mycobacterium tuberculosis infection: a systematic review and meta-analysis (vol. 37, pg. 88, 2011)", Eur. Respir. J, 39, pp.793~793 (2012)
- Guenat S, "Homogeneous and nonradioactive high-throughput screening platform for the characterization of kinase inhibitors in cell lysates", J. Biomol. Screen, 11, pp.1015~1026 (2006)
- Lawn SD, "Tuberculosis", Lancet, 378, pp.57~72 (2011)
- Lee J, "Comparison of two commercial interferon-γ assays for diagnosing Mycobacterium tuberculosis infection", Eur. Respir. J, 28, pp.24~30 (2006)
- Mandalakas AM, "Interferon-gamma release assays and childhood tuberculosis: systematic review and meta-analysis", Int. J. Tuberc. Lung Dis, 15, pp.1018~1032 (2011)
- Mazurek GH, "Guidelines for using the QuantiFERON (R)-TB gold test for detecting Mycobacterium tuberculosis infection, United States. MMWR Morbid", Mortal. Wkly. Rep, 54, pp.49~55 (2005)
- Menzies D, "Meta-analysis: new tests for the diagnosis of latent tuberculosis infection: areas of uncertainty and recommendations for research", Ann. Int. Med, 146, pp.340~354 (2007)
- Monneret D, "Evaluation of LOCI® technology-based thyroid blood tests on the Dimension Vista® analyzer", Clin. Biochem, 46, pp.1290~1297 (2013)
- Montoya D, "IL-32 is a molecular marker of a host defense network in human tuberculosis", Sci. Transl. Med, 6, pp.250ra114~ (2014)
- Pai M, "Gamma interferon release assays for detection of Mycobacterium tuberculosis infection", Clin. Microbiol. Rev, 27, pp.3~20 (2014)
- Pai M, "Systematic review: Tcell-based assays for the diagnosis of latent tuberculosis infection: an update", Ann. Int. Med, 149, pp.177~184 (2008)
- Santin M, "Interferon-gamma release assays for the diagnosis of tuberculosis and tuberculosis infection in HIV-infected adults: a systematic review and meta-analysis", PLoS One, pp.7~ (2012)
- Villarino ME, "Prevention and treatment of tuberculosis among patients infected with human immunodeficiency virus: principles of therapy and revised recommendations", MMWR Morbid. Mortal. Wkly. Rep, pp.47~ (1998)
- Global Tuberculosis Report 2013, "WHO", World Health Organization, ~ (2013)
- Global Tuberculosis Report 2014, "WHO", World Health Organization, ~ (2014)