Graphically encoded suspension array for multiplex immunoassay and quantification of autoimmune biomarkers in patient sera

To provide a high-throughput, efficient, and accurate method to monitor multiple-fungicide resistance of Botrytis cinerea in the field, we used the suspension array, sequencing, and mycelial growth assay in our research. Discriminating-dose bioassays for detecting carbendazim, diethofencarb, boscalid, and iprodione resistance (CarR, DieR, BosR, and IprR, respectively) were used to analyze 257 isolates collected from Hebei Province in China during 2016 and 2017. High resistance frequencies to carbendazim (100%), diethofencarb (92.08%), and iprodione (86.59%) were detected. BosR isolates accounted for 11.67% of the total. In addition, 103 isolates were randomly selected for phenotype and genotype detection. The high-throughput suspension array was utilized to detect eight genotypes simultaneously, including BenA-E198, BenA-198A, SdhB-H272, SdhB-272Y, BcOS1-I365, BcOS1-365S, erg27-F412, and erg27-412S, which were associated with resistance toward carbendazim or diethofencarb, boscalid, iprodione, and fenhexamid (FenR), respectively. Most of the benzimidazole-resistant isolates (81.55%) possessed the E198V mutation in the BenA gene. Ninety-three isolates with dual resistance to carbendazim and diethofencarb showed the E198V/K mutation. All BosR isolates carried the H272R mutation in the SdhB gene. The I365S and Q369P+N373S (66.99%) mutations in the BcOS1 gene were more frequently observed. No mutation was detected in the erg27 gene in Hebei isolates. There were 13 resistance profile phenotypes. Phenotypes with triple resistance were the most common (83.50%), and CarRDieRBosSIprRFenS was the major type. CarR isolates that carried E198V/K/A were all highly resistant (HR) and only one F200Y mutant was moderately resistant (MR) to carbendazim. Isolates that possessed E198V/K were MR or HR to diethofencarb. BosR isolates that possessed H272R mutation were lowly resistant (LR). IprR isolates were all LR or MR. The distribution of half maximal effective concentrations of CarR isolates with E198V/K mutations and IprR isolates with Q369P+N373S mutations significantly increased from 2016 to 2017. Combined with our observations, a combination method of the high-throughput suspension array and the mycelial growth assay was suggested to accurately monitor multiple resistance of B. cinerea in the field.

Download Full-text

Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays

Malaria Journal ◽

10.1186/1475-2875-10-192 ◽

2011 ◽

Vol 10 (1) ◽

pp. 192 ◽

Cited By ~ 24

Author(s):

Edmilson Rui ◽

Carmen Fernandez-Becerra ◽

Satoru Takeo ◽

Sergi Sanz ◽

Marcus VG Lacerda ◽

...

Keyword(s):

Escherichia Coli ◽

Plasmodium Vivax ◽

Wheat Germ ◽

Suspension Array

Download Full-text

A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses

BioMed Research International ◽

10.1155/2015/678084 ◽

2015 ◽

Vol 2015 ◽

pp. 1-13 ◽

Cited By ~ 29

Author(s):

Cécile Beck ◽

Philippe Desprès ◽

Sylvie Paulous ◽

Jessica Vanhomwegen ◽

Steeve Lowenski ◽

...

Keyword(s):

High Performance ◽

Neurological Diseases ◽

Expression System ◽

Cross Reactivity ◽

Encephalitis Virus ◽

Serological Tests ◽

Multiplex Immunoassay ◽

Tick Borne Encephalitis ◽

Tick Borne Encephalitis Virus ◽

Negative Controls

West Nile virus (WNV), Japanese encephalitis virus (JEV), and tick-borne encephalitis virus (TBEV) are flaviviruses responsible for severe neuroinvasive infections in humans and horses. The confirmation of flavivirus infections is mostly based on rapid serological tests such as enzyme-linked immunosorbent assays (ELISAs). These tests suffer from poor specificity, mainly due to antigenic cross-reactivity among flavivirus members. Robust diagnosis therefore needs to be validated through virus neutralisation tests (VNTs) which are time-consuming and require BSL3 facilities. The flavivirus envelope (E) glycoprotein ectodomain is composed of three domains (D) named DI, DII, and DIII, with EDIII containing virus-specific epitopes. In order to improve the serological differentiation of flavivirus infections, the recombinant soluble ectodomain of WNV E (WNV.sE) and EDIIIs (rEDIIIs) of WNV, JEV, and TBEV were synthesised using theDrosophilaS2 expression system. Purified antigens were covalently bonded to fluorescent beads. The microspheres coupled to WNV.sE or rEDIIIs were assayed with about 300 equine immune sera from natural and experimental flavivirus infections and 172 nonimmune equine sera as negative controls. rEDIII-coupled microspheres captured specific antibodies against WNV, TBEV, or JEV in positive horse sera. This innovative multiplex immunoassay is a powerful alternative to ELISAs and VNTs for veterinary diagnosis of flavivirus-related diseases.

Download Full-text

A Single Snapshot Multiplex Immunoassay Platform Utilizing Dense Test Lines Based on Engineered Beads

Biosensors and Bioelectronics ◽

10.1016/j.bios.2021.113388 ◽

2021 ◽

pp. 113388

Author(s):

Wonhyung Lee ◽

Hojin Kim ◽

Pan Kee Bae ◽

Sanghyun Lee ◽

Sung Yang ◽

...

Keyword(s):

Multiplex Immunoassay ◽

Single Snapshot

Download Full-text

Abstract 4336: Interrogating the Human Myocardial Interstitium during Myocardial Arrest and Reperfusion: Dynamic Changes in Cytokines and Protease Activity

Circulation ◽

10.1161/circ.118.suppl_18.s_867-a ◽

2008 ◽

Vol 118 (suppl_18) ◽

Author(s):

Robert E Stroud ◽

Christine N Koval ◽

Isabelle Gengler ◽

Anne M Deschamps ◽

John S Ikonomidis ◽

...

Keyword(s):

Detection System ◽

Ischemia Reperfusion ◽

Dynamic Assessment ◽

High Sensitivity ◽

Fluorescent Detection ◽

Suspension Array ◽

Cytokine Analysis ◽

And Function ◽

Inflammatory Molecules ◽

Pulmonary Bypass

Background. Cytokines, such as the interleukins (IL1β, IL2, IL6) and tumor necrosis factor (TNF) can modulate myocardial structure and function with ischemia/reperfusion (I/R) but dynamic assessment of these biological molecules within the human myocardial interstitium with I/R has not been performed, and the inter-relationship to matrix metalloproteinases activity (MMPact) remains unexplored. Accordingly, a fluorogenic microdialysis method was used to simultaneously measure myocardial interstitial cytokine levels and MMPact in patients during and following I/R. Methods . MMPact was measured in patients (n=13) undergoing cardio-pulmonary bypass (CPB) at baseline, during myocardial arrest and CPB (on-CPB), and immediately following reperfusion and separation from CPB (post-CPB) by a validated in-line microdialysis fluorescent detection system. Myocardial interstitial fluid was subjected to cytokine analysis by high sensitivity multiplex suspension array. Results . Interstitial MMPact increased by over 30% post-CPB and was accompanied by a specific change in cytokine profiles (Figure ). The classical pro-inflammatory molecules such as TNF and IL6 were either not detectable or unchanged, whereas IL1β and IL2 which can be proinflammatory, were increased. Conclusions. These unique results demonstrated that a dynamic cytokine signature occurs within the human myocardial interstitium following I/R and is temporally related to heightened MMP activity. Direct interrogation of the human myocardial interstitium may provide a unique insight into critical signaling pathways which may evoke adverse structural and functional events following I/R.

Download Full-text

Abstract 3050: Assessment of Genetic Risk for Thoracic Aortic Aneurysm Among Hypertensive Patients

Circulation ◽

10.1161/circ.116.suppl_16.ii_683-b ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Kimihiko Kato ◽

Mitsutoshi Oguri ◽

Tetsuro Yoshida ◽

Takeshi Hibino ◽

Kazuhiro Yajima ◽

...

Keyword(s):

Risk Factor ◽

Genetic Risk ◽

Odds Ratio ◽

The United States ◽

Multivariable Logistic Regression Analysis ◽

Forward Selection ◽

Structural Formation ◽

Hypertensive Patients ◽

Mortality And Morbidity ◽

Suspension Array

Introduction: Aneurysms and dissections at the thoracic aorta (TAA) are the leading causes of mortality and morbidity in the United States and Japan. Although hypertension is a major risk for TAA, several studies have indicated that genetic factors also influence the structural formation of TAA. The purpose of the present study was to identify gene polymorphisms associated with TAA among hypertensive patients for assessment of the genetic risk for this condition. Methods: This case-control study comprised 1351 hypertensive individuals, including 88 subjects with TAA and 1263 without any cardiovascular complications. The genotypes for 142 polymorphisms of 121 candidate genes were determined by a method that combines the polymerase chain reaction and sequence-specific oligonucleotide probes with suspension array technology. Results: Evaluation of genotype distributions by the chi-square test and subsequent multivariable logistic regression analysis with adjustment for age, sex, body mass index, and prevalence of smoking, diabetes mellitus, hypercholesterolemia, and metabolic syndrome revealed that the T→G (3′ UTR) polymorphism of the thrombospondin 2 gene ( THBS2 ; odds ratio, 4.7) is significantly (P < 0.05) associated with TAA in hypertensive subjects, with the variant G allele representing a risk factor for this condition. Furthermore, the -110A→C polymorphism of heat-shock 70-kD protein 8 gene ( HSPA8 ; odd ratio, 0.3), the C→T (Pro198Leu) polymorphism of the glutathione peroxidase gene ( GPX1 ; odds ratio, 0.3), the -6G→A polymorphism of angiotensin 1 gene ( AGT ; odds ratio, 0.3), and the -850C→T polymorphism of tumor necrosis factor gene ( TNF ; odds ratio, 0.5) were significantly associated with this condition, with the variant allele of each polymorphism being protective against this condition. A stepwise forward selection procedure revealed that polymorphisms of THBS2 , HSPA8 , GPX1 , AGT , and TNF significantly affected the prevalence of TAA. Conclusions: Genetic variant of THBS2 may be a risk factor for TAA in hypertensive patients, whereas variants of HSPA8 , GPX1 , AGT , and TNF may be protective against this condition. Determination of genotypes for these polymorphisms may prove informative for assessment of the genetic risk for TAA.

Download Full-text