Validation of multiplex microbead immunoassay for simultaneous serodetection of multiple infectious agents in laboratory mouse

Ravindran, Resmi; Khan, Imran; Krishnan, Viswanathan V; Ziman, Melanie; Kendall, Lon V.; Frasier, Janelle M.; Bates, R. W.; Griffey, Steve M.; Fahey, James R.; Luciw, Paul A.

doi:10.1016/j.jim.2010.10.003

Cited by 19 publications

(40 citation statements)

References 18 publications

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“…Others have validated multiplex assays of hormones (Sun et al, 2010) and infectious agents (Ravindran et al, 2010) in mouse plasma but not cytokines. We show that a mouse MSD 7-plex cytokine immunoassay has intra-and inter-assay CV% <30% for the majority of analytes and percent recovery of spiked samples in both plasma (4 of 7 cytokines with mean % recovery 75-125%) and BAL (5 of 7 cytokines with mean % recovery 75-125%) ( Figure 6 ).…”

Section: Discussionmentioning

confidence: 99%

Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples

Bastarache

Koyama

Wickersham

et al. 2014

Journal of Immunological Methods

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Despite the widespread use of multiplex immunoassays, there are very few scientific reports that test the accuracy and reliability of a platform prior to publication of experimental data. Our laboratory has previously demonstrated the need for new assay platform validation prior to use of biologic samples from large studies in order to optimize sample handling and assay performance. In this study, our goal was to test the accuracy and reproducibility of an electrochemiluminescent multiplex immunoassay platform (Meso Scale Discovery, MSD®) and compare this platform to validated, singleplex immunoassays (R&D Systems®) using actual study subject (human plasma and mouse bronchoalveolar lavage fluid (BALF) and plasma) samples. We found that the MSD platform performed well on intra- and inter-assay comparisons, spike and recovery and cross-platform comparisons. The mean intra-assay CV% and range for MSD was 3.49 (0.0-10.4) for IL-6 and 2.04 (0.1-7.9) for IL-8. The correlation between values for identical samples measured on both MSD and R&D was R=0.97 for both analytes. The mouse MSD assay had a broader range of CV% with means ranging from 9.5-28.5 depending on the analyte. The range of mean CV% was similar for single plex ELISAs at 4.3-23.7 depending on the analyte. Regardless of species or sample type, CV% was more variable at lower protein concentrations. In conclusion, we validated a multiplex electrochemiluminscent assay system and found that it has superior test characteristics in human plasma compared to mouse BALF and plasma. Both human and MSD assays compared favorably to well-validated singleplex ELISA's

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Section: Discussionmentioning

confidence: 99%

Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples

Bastarache

Koyama

Wickersham

et al. 2014

Journal of Immunological Methods

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“…In contrast, the regression-based classification algorithms used in this study provided a powerful summary of prediction and performance in the presence of multiple positive signals. In the development of a computational data-mining system, the above-described algorithms provided the most accurate results with data from several hundred samples (unpublished data) analyzed by multiplex serodetection (10 infectious pathogens) in a validated mouse model of infectious diseases (20). Anti-M. tuberculosis antibody profiles.…”

Section: Discussionmentioning

confidence: 99%

“…Cluster analysis was performed to define natural groupings of antigens and TB patients (AFB ϩ group) analyzed with respect to healthy controls (IH group) as previously described (20). Briefly, data sets with which to perform the cluster analysis were generated by scaling the MFI levels of each sample in the AFB ϩ group with reference to the baseline value for each microbead set.…”

Section: Methodsmentioning

confidence: 99%

Plasma Antibody Profiles as Diagnostic Biomarkers for Tuberculosis

Khan

Ravindran

Krishnan

et al. 2011

Clin Vaccine Immunol

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Two billion people are infected with Mycobacterium tuberculosis, the etiological agent of tuberculosis (TB), worldwide. Ten million to 20 million of the infected individuals develop disease per year. TB is a treatable disease, provided that it is diagnosed in a timely manner. The current TB diagnostic methods are subjective, inefficient, or not cost-effective. Antibody-based blood tests can be used efficiently and cost-effectively for TB diagnosis. A major challenge is that different TB patients generate antibodies against different antigens. Therefore, a multiplex immunoassay approach is needed. We have developed a multiplex panel of 28 M. tuberculosis antigen-coated microbeads. Plasma samples were obtained from over 300 pulmonary TB patients and healthy controls in a country where TB is endemic, Pakistan. Multiplex data were analyzed using computational tools by multivariate statistics, classification algorithms, and cluster analysis. The results of antibody profile-based detection, using 16 selected antigens, closely correlated with those of the sputum-based diagnostic methods (smear microscopy and culture) practiced in countries where TB is endemic. Multiplex microbead immunoassay had a sensitivity and specificity of approximately 90% and 80%, respectively. These antibody profiles could potentially be useful for the diagnosis of nonpulmonary TB, which accounts for approximately 20% of cases of disease. Since an automated, high-throughput version of this multiplex microbead immunoassay could analyze thousands of samples per day, it may be useful for the diagnosis of TB in millions of patients worldwide.More than one-third of the world's population is infected with Mycobacterium tuberculosis (7, 26a). Annually, 10 million to 20 million of these individuals develop clinical symptoms, and about 2 million die of tuberculosis (TB) (4, 17a). The infected host typically mounts a vigorous immune response (25). Nevertheless, 10% of all infections result in active disease within 2 years. Another 10% of cases may experience disease after a latent phase spanning many years (8, 17a). Several Mycobacterium species (e.g., M. tuberculosis, M. bovis, and M. africanum) can infect and cause disease in humans (2, 24). In about 80% of active TB cases, direct involvement of the lung results in pulmonary disease (4a). However, M. tuberculosis can spread to other organs. In approximately 20% of cases, M. tuberculosis may cause nonpulmonary disease in various organ systems (urogenital system, nervous system, digestive system, skeletal system, etc.) with or without the lung involvement (7,18). TB is a treatable disease, provided that a timely and appropriate diagnosis is made (4a). Commonly used sputumbased methods for pulmonary TB diagnosis are subjective, insensitive, and/or inefficient. Furthermore, for the detection of pediatric pulmonary TB, a major limitation is that children often have difficulty producing usable quantities of sputum.Sputum smear acid-fast bacillus (AFB) microscopy is recommended by the World Health Organizatio...

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“…These proteins are critical regulators of normal immune responses, inflammatory reactions and cell proliferation, and play key roles in chronic conditions such as heart disease, diabetes, and cancer, as well as acute conditions, such as infections (8)(9)(10)(11)(12)(13)(14).…”

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confidence: 99%

Multiplexed measurements of immunomodulator levels in peripheral blood of healthy subjects: Effects of analytical variables based on anticoagulants, age and gender

et al. 2013

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Validation of multiplex microbead immunoassay for simultaneous serodetection of multiple infectious agents in laboratory mouse

Cited by 19 publications

References 18 publications

Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples

Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples

Plasma Antibody Profiles as Diagnostic Biomarkers for Tuberculosis

Multiplexed measurements of immunomodulator levels in peripheral blood of healthy subjects: Effects of analytical variables based on anticoagulants, age and gender

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