Novel immunoassay for diagnosis of ongoing Clostridioides difficile infections using serum and medium enriched for newly synthesized antibodies (MENSA)

Haddad, Natalie S.; Nozick, Sophia; Kim, Geena; Ohanian, Shant; Kraft, Colleen S.; Rebolledo, Paulina A.; Wang, Yun; Wu, Hao; Bressler, Adam; Shi, Le; Kuruvilla, Merin; Cannon, Leah; Lee, F. Eun-Hyung; Daiss, John L.

doi:10.1016/j.jim.2020.112932

Cited by 9 publications

(21 citation statements)

References 77 publications

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“…After approval from Emory University, Dekalb (now Emory Dekalb) and Grady Institutional Review Boards, patients with primary CDI were enrolled and followed for recurrence. The primary CDI population (n=46) was similar in size to the control population (n=64) and in the proportion of races ( p =0.52) and sexes ( p =0.21; Table 1) (25). The control population was younger than the CDI population ( p <0.0001), but there was no significant difference between younger and older healthy controls in serum and MENSA levels of antibodies specific for C. difficile antigens (MENSA p =0.71; Serum p=0.07, older subjects slightly lower).…”

Section: Resultsmentioning

confidence: 96%

“…For this test, of the C. difficile antigens, the secreted toxins, TcdB-CROP, TcdBvir-CROP and CDTb provided the best results. Although C. difficile displays an array of cell surface and secreted antigens and extensively described in our previous studies (25), a defensive enzymes (GDH (34)), a flagellar components (FliC (35-37)) and a cell wall protein associated with biofilm building (Cwp84 (38-41)) added little or no value for MENSA or serum-based diagnostics. Antibody levels specific for the TcdA domains, CROP and GTD, were redundant with the larger responses to the TcdB and CDTb toxins.…”

Section: Discussionmentioning

confidence: 97%

“…Selection and design of CD recombinant antigens are presented in detail (25). Briefly, antigens or domains thereof were selected based on known immunogenicity and/or pathogenicity.…”

Section: Methodsmentioning

confidence: 99%

“…All samples were tested using a multiplex immunoassay protocol previously described using Luminex MAGPIX® (25). All MENSA samples were tested undiluted; serum samples were diluted 1:1000 in Assay Buffer (PBS, 1% BSA, pH7.5).…”

Section: Methodsmentioning

confidence: 99%

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Antibodies in Serum and MENSA Predict Non-recurrence in Primary Clostridioides difficile infection

Haddad¹,

Nozick²,

Kim³

et al. 2021

Preprint

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BackgroundWithin eight weeks of primary Clostridioides difficile infection (CDI), as many as 30% of patients develop recurrent disease with the associated risks of multiple relapses, morbidity, and economic burden. There are no validated biomarkers predictive of recurrence during primary infection. This study demonstrates the potential of a simple test for identifying hospitalized CDI patients at low risk for disease recurrence.MethodsForty-six hospitalized CDI patients were enrolled at Emory University Hospitals. Serum and MENSA samples prepared during weeks 1, 2, and 4 following symptom-onset were measured for antibodies specific for ten C. difficile antigens.ResultsAmong the 46 C. difficile-infected patients, nine (19.5%) experienced recurrence within eight weeks of primary infection. Among the 37 non-recurrent patients, 23 had anti-C. difficile MENSA antibodies specific for any of the three toxin antigens: TcdB-CROP, TcdBvir-CROP, and/or CDTb. Positive MENSA responses occurred within the first week post-symptom onset, including six patients who never seroconverted. A similar trend was observed in serum responses, but they peaked later and identified fewer patients (19/37). In contrast, none of the patients who subsequently recurred after hospitalization produced antibodies specific for the three C. difficile toxin antigens. IgA antibodies for the toxin antigens demonstrated the greatest predictive power for protection from recurrence.DiscussionThe development of IgG and/or IgA antibodies for three C. difficile toxins in serum and/or MENSA has prognostic potential. These immunoassays measure nascent immune responses that reduce the likelihood of recurrence. Early identification of patients at-risk for recurrence can reduce costs and morbidity.

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Section: Resultsmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 97%

“…Selection and design of CD recombinant antigens are presented in detail (25). Briefly, antigens or domains thereof were selected based on known immunogenicity and/or pathogenicity.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Antibodies in Serum and MENSA Predict Non-recurrence in Primary Clostridioides difficile infection

Haddad¹,

Nozick²,

Kim³

et al. 2021

Preprint

Self Cite

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“…ASC appear as early as two days post-symptom onset (DPSO) and disappear when the infection has resolved [17]. These attributes have sparked interest in the development of assays using pathogen-specific ASC for diagnosis of active infections [18][19][20]. Direct enumeration of ASC by ELISpot assays or measurement of antibodies from circulating ASC collected in vitro in a novel matrix, MENSA (media enriched with newly synthesized antibodies), have been shown to be effective diagnostics [16][17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Diagnosis of Streptococcus pneumoniae infection using circulating antibody secreting cells

et al. 2021

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Background Streptococcus pneumoniae infections cause morbidity and mortality worldwide. A rapid, simple diagnostic method could reduce the time needed to introduce definitive therapy potentially improving patient outcomes. Methods We introduce two new methods for diagnosing S. pneumoniae infections by measuring the presence of newly activated, pathogen-specific, circulating Antibody Secreting Cells (ASC). First, ASC were detected by ELISpot assays that measure cells secreting antibodies specific for signature antigens. Second, the antibodies secreted by isolated ASC were collected in vitro in a novel matrix, MENSA (media enriched with newly synthesized antibodies) and antibodies against S. pneumoniae antigens were measured using Luminex immunoassays. Each assay was evaluated using blood from S. pneumoniae and non-S. pneumoniae-infected adult patients. Results We enrolled 23 patients with culture-confirmed S. pneumoniae infections and 24 controls consisting of 12 non-S. pneumoniae infections, 10 healthy donors and two colonized with S. pneumoniae. By ELISpot assays, twenty-one of 23 infected patients were positive, and all 24 controls were negative. Using MENSA samples, four of five S. pneumoniae-infected patients were positive by Luminex immunoassays while all five non-S. pneumoniae-infected patients were negative. Conclusion Specific antibodies produced by activated ASC may provide a simple diagnostic for ongoing S. pneumoniae infections. This method has the potential to diagnose acute bacterial infections.

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StaphAIR: A Label-Free Antigen Microarray Approach to Detecting Anti-Staphylococcus aureus Antibody Responses in Orthopedic Infections

Klose

Daiss

et al. 2021

Anal. Chem.

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Arrayed imaging reflectometry (AIR) is an optical biosensor platform for simple, multiplex measurement of antigenspecific antibody responses in patient blood samples. Here, we report the development of StaphAIR, an 8-plex Staphylococcus aureus antigen array on the AIR platform for profiling antigenspecific anti-S. aureus humoral immune responses. Initial validation experiments with mouse and humanized monoclonal antibodies against the S. aureus autolysin glucosaminidase (Gmd) domain, and subsequent testing with dilution series of pooled positive human serum confirmed analytically robust behavior of the array, with all antigens displaying Langmuir-type dose−response curves. Testing a cohort of 82 patients with S. aureus musculoskeletal infections (MSKI) and 30 healthy individuals enabled discrimination of individual patient responses to different S. aureus antigens, with statistical significance between osteomyelitis patients and controls obtained overall for four individual antigens (IsdA, IsdB, Gmd, and SCIN). Multivariate analyses of the antibody titers obtained from StaphAIR revealed its utility as a potential diagnostic tool for detecting S. aureus MSKI (area under the receiver operating characteristic curve (AUC) > 0.85). We conclude that StaphAIR has utility as a high-throughput immunoassay for studying and diagnosing osteomyelitis in patients.

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Novel immunoassay for diagnosis of ongoing Clostridioides difficile infections using serum and medium enriched for newly synthesized antibodies (MENSA)

Cited by 9 publications

References 77 publications

Antibodies in Serum and MENSA Predict Non-recurrence in Primary Clostridioides difficile infection

Antibodies in Serum and MENSA Predict Non-recurrence in Primary Clostridioides difficile infection

Diagnosis of Streptococcus pneumoniae infection using circulating antibody secreting cells

StaphAIR: A Label-Free Antigen Microarray Approach to Detecting Anti-Staphylococcus aureus Antibody Responses in Orthopedic Infections

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