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Zeptomole per milliliter detection and quantification of edema factor in plasma by LC-MS/MS yields insights into toxemia and the progression of inhalation anthrax.

التفاصيل البيبلوغرافية
العنوان: Zeptomole per milliliter detection and quantification of edema factor in plasma by LC-MS/MS yields insights into toxemia and the progression of inhalation anthrax.
المؤلفون: Lins RC; Battelle Atlanta Analytical Services, 2987 Clairmont Road NE, Suite 450, Atlanta, GA, 30329, USA., Boyer AE; Centers for Disease Control and Prevention, 4770 Buford Highway Mailstop F-50, Atlanta, GA, 30341, USA. aboyer@cdc.gov., Kuklenyik Z; Centers for Disease Control and Prevention, 4770 Buford Highway Mailstop F-50, Atlanta, GA, 30341, USA., Woolfitt AR; Centers for Disease Control and Prevention, 4770 Buford Highway Mailstop F-50, Atlanta, GA, 30341, USA., Goldstein J; Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30333, USA., Hoffmaster AR; Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30333, USA., Gallegos-Candela M; Centers for Disease Control and Prevention, 4770 Buford Highway Mailstop F-50, Atlanta, GA, 30341, USA., Leysath CE; Laboratory of Parasitic Diseases, NIAID, NIH, 9000 Rockville Pike, Bethesda, MD, 20892, USA.; Center for Innovation in Advanced Development and Manufacturing, Texas A&M University Health Science Center, 8441 Riverside Parkway, Suite 3200, Bryan, TX, 77807, USA., Chen Z; Laboratory of Infectious Diseases, National Institute for Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), 9000 Rockville Pike, Bethesda, MD, 20892, USA., Brumlow JO; Battelle Atlanta Analytical Services, 2987 Clairmont Road NE, Suite 450, Atlanta, GA, 30329, USA., Quinn CP; Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30333, USA., Bagarozzi DA Jr; Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30333, USA., Leppla SH; Laboratory of Parasitic Diseases, NIAID, NIH, 9000 Rockville Pike, Bethesda, MD, 20892, USA., Barr JR; Centers for Disease Control and Prevention, 4770 Buford Highway Mailstop F-50, Atlanta, GA, 30341, USA.
المصدر: Analytical and bioanalytical chemistry [Anal Bioanal Chem] 2019 May; Vol. 411 (12), pp. 2493-2509. Date of Electronic Publication: 2019 Mar 26.
نوع المنشور: Journal Article; Validation Study
اللغة: English
بيانات الدورية: Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 101134327 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1618-2650 (Electronic) Linking ISSN: 16182642 NLM ISO Abbreviation: Anal Bioanal Chem Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Heidelberg : Springer-Verlag, 2002-
مواضيع طبية MeSH: Anthrax/*pathology , Antigens, Bacterial/*blood , Bacillus anthracis/*pathogenicity , Bacterial Toxins/*blood , Chromatography, High Pressure Liquid/*methods , Respiratory Tract Infections/*pathology , Tandem Mass Spectrometry/*methods , Toxemia/*pathology, Adenosine Triphosphate/metabolism ; Animals ; Anthrax/blood ; Case-Control Studies ; Cyclic AMP/biosynthesis ; Disease Progression ; Enzyme-Linked Immunosorbent Assay ; Humans ; Limit of Detection ; Macaca mulatta ; Polymerase Chain Reaction ; Respiratory Tract Infections/blood ; Toxemia/blood ; Toxemia/microbiology
مستخلص: Inhalation of Bacillus anthracis spores can cause a rapidly progressing fatal infection. B. anthracis secretes three protein toxins: lethal factor (LF), edema factor (EF), and protective antigen (PA). EF and LF may circulate as free or PA-bound forms. Both free EF (EF) and PA-bound-EF (ETx) have adenylyl cyclase activity converting ATP to cAMP. We developed an adenylyl cyclase activity-based method for detecting and quantifying total EF (EF+ETx) in plasma. The three-step method includes magnetic immunocapture with monoclonal antibodies, reaction with ATP generating cAMP, and quantification of cAMP by isotope-dilution HPLC-MS/MS. Total EF was quantified from 5PL regression of cAMP vs ETx concentration. The detection limit was 20 fg/mL (225 zeptomoles/mL for the 89 kDa protein). Relative standard deviations for controls with 0.3, 6.0, and 90 pg/mL were 11.7-16.6% with 91.2-99.5% accuracy. The method demonstrated 100% specificity in 238 human serum/plasma samples collected from unexposed healthy individuals, and 100% sensitivity in samples from 3 human and 5 rhesus macaques with inhalation anthrax. Analysis of EF in the rhesus macaques showed that it was detected earlier post-exposure than B. anthracis by culture and PCR. Similar to LF, the kinetics of EF over the course of infection were triphasic, with an initial rise (phase-1), decline (phase-2), and final rapid rise (phase-3). EF levels were ~ 2-4 orders of magnitude lower than LF during phase-1 and phase-2 and only ~ 6-fold lower at death/euthanasia. Analysis of EF improves early diagnosis and adds to our understanding of anthrax toxemia throughout infection. The LF/EF ratio may also indicate the stage of infection and need for advanced treatments.
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معلومات مُعتمدة: CC999999 United States ImCDC Intramural CDC HHS
فهرسة مساهمة: Keywords: Adenylyl cyclase; Anthrax; Cyclic AMP; Edema toxin; HPLC; Mass spectrometry
المشرفين على المادة: 0 (Antigens, Bacterial)
0 (Bacterial Toxins)
0 (anthrax toxin)
8L70Q75FXE (Adenosine Triphosphate)
E0399OZS9N (Cyclic AMP)
SCR Disease Name: Inhalation anthrax
تواريخ الأحداث: Date Created: 20190327 Date Completed: 20190624 Latest Revision: 20210109
رمز التحديث: 20221213
مُعرف محوري في PubMed: PMC6988385
DOI: 10.1007/s00216-019-01730-4
PMID: 30911800
قاعدة البيانات: MEDLINE
الوصف
تدمد:1618-2650
DOI:10.1007/s00216-019-01730-4