scholarly journals Development of a Nucleocapsid-Based Human Coronavirus Immunoassay and Estimates of Individuals Exposed to Coronavirus in a U.S. Metropolitan Population

2008 ◽  
Vol 15 (12) ◽  
pp. 1805-1810 ◽  
Author(s):  
Emily G. Severance ◽  
Ioannis Bossis ◽  
Faith B. Dickerson ◽  
Cassie R. Stallings ◽  
Andrea E. Origoni ◽  
...  

ABSTRACT Coronaviruses cause respiratory infections ranging from common colds to severe acute respiratory syndrome (SARS) in humans. Estimates for exposure to non-SARS coronaviruses are high, particularly for 229E and OC43; however, less information regarding seroprevalence is available for HKU1 and NL63. To measure exposure rates to these four coronavirus strains (229E, HKU1, NL63, and OC43), we devised an immunoassay based on amino- and carboxy-terminally tagged recombinant coronavirus nucleocapsid antigens. Four human and one feline coronavirus antigen were cloned into baculoviruses expressed in insect cells and recovered proteins bound in the solid phase of an enzyme-linked immunosorbent assay-based system. We screened sera from 10 children and 196 adults and established primary cutoff points based on immunoglobulin G (IgG) antibody levels of the predominantly seronegative children. The proportion of seropositive adults for each coronavirus was as follows: 229E, 91.3%; HKU1, 59.2%; NL63, 91.8%; and OC43, 90.8%. No evidence of a significant serological response to the feline coronavirus was observed. Significant associations of coronavirus seropositivity and antibody levels with age, gender, race, socioeconomic status, smoking status, and season of the blood draw were tested with chi-square and regression analyses. The group II coronaviruses (OC43 and HKU1) were significantly associated with race (P ≤ 0.009 and P ≤ 0.03, respectively). Elevated OC43 IgG levels were further significantly associated with smoking status (P ≤ 0.03), as were high NL63 titers with socioeconomic status (P ≤ 0.04). The high-level immunoreactivity of each coronavirus was significantly associated with the summer season (P ≤ 0.01 to 0.0001). In summary, high rates of exposure to 229E, NL63, and OC43 and a moderate rate of exposure to HKU1 characterized the seroprevalence among individuals in this population. Demographic factors, such as race, smoking status, and socioeconomic status, may confer an increased risk of susceptibility to these viruses.

Author(s):  
Omid Nekouei ◽  
Paolo Martelli ◽  
Sophie St-Hilaire ◽  
Hui Suk Wai ◽  
Karthiyani Krishnasamy ◽  
...  

Rabies is a fatal zoonotic disease that can affect all mammals. Following the directives of the rabies ordinance of the Government of Hong Kong, all wild macaques captured under an ongoing sterilization program (since 2000) were vaccinated against rabies. The main objective of this study was to assess the serological response to rabies vaccination in the population of Hong Kong macaques. An inactivated rabies vaccine was subcutaneously administered to captured macaques under anesthesia. In a 2015 field survey, blood samples from the animals were collected and stored in -80℃ freezer. In July 2021, all frozen sera from vaccinated animals were prepared and tested for antibodies against rabies virus using a commercial enzyme-linked immunosorbent assay (ELISA) test. The test results were dichotomized at the recommended cut-off point of the test kit. Sixty-five samples from the vaccinated macaques were available for this study. All of these animals had received at least one dose of vaccine (1 vaccination) between 2008 and 2015. The interval between the 1 vaccination and blood sampling dates ranged from 21 to 2,779 days. Only five of the 65 macaques had a second vaccination record at the time of sampling; all five had high antibody levels. Among the remaining macaques, 77% (46/60) were positive for rabies antibodies. No specific association was observed between the post-vaccination period and the antibody titer of these macaques and no adverse reactions to vaccination were reported. The current vaccination strategy in Hong Kong macaques appears to effectively elicit rabies antibodies in a high proportion of macaque populations in the wild (78-87%). However, reaching the precise level of protection against a potential challenge with the virus should further be investigated.


2020 ◽  
Vol 38 (15_suppl) ◽  
pp. 12062-12062
Author(s):  
Leslie Ann Modlin ◽  
N. Ari Wijetunga ◽  
Minal Patel ◽  
Teng Gao ◽  
Ryan Ptashkin ◽  
...  

12062 Background: Clonal hematopoiesis (CH), characterized by recurrent somatic mutations in blood, is a common age-associated condition that portends an increased risk of myeloid neoplasms and cardiac disease. Oncologic therapies appear to promote CH, including ionizing radiation therapy (RT) (OR = 1.4, p < 10−6) and systemic DNA-damaging agents (OR = 1.2, p = 8x10−4). How various RT parameters (e.g. target site, dose, fractionation, modality) may influence CH is unknown. Methods: CH mutations were identified via targeted, deep-coverage next-generation sequencing from paired peripheral blood and tumor samples (MSK-IMPACT). CH was defined as a somatic blood mutation with a minimum variant allele frequency of 2%. Putative driver mutations (CH-PD) were identified from OncoKB and other published sources. Clinical and RT characteristics were abstracted from medical records. To account for differences in RT dose and fractionation, equivalent radiation dose in 2 Gy fractions (EQD2) with an α/β ratio of 3 for late effects was calculated. Univariate and logistic regression modeling for associations between clinical and treatment parameters and CH were performed. Results: We identified 2,195 patients who received RT before blood draw and 7,832 who did not, encompassing 57 histologies. A median of 267 days elapsed between the end of RT and blood draw. After RT, 22% of patients had at least one CH-PD mutation (n = 486). The most common single anatomic sites radiated were pelvis, chest wall/breast, and head and neck. Conventional RT was used in 2% (n = 46), 3D-conformal in 14% (n = 308), intensity modulated RT in 36% (n = 787), volumetric modulated arc RT in 12% (n = 263), multiple techniques in 26% (n = 560), and unknown in 11% (n = 231). There was no association between RT modality and presence of CH-PD (p > 0.05 for all between group comparisons of modality). On multivariate regression after controlling for age, race, time from diagnosis to blood draw, smoking status, and for chemotherapy class, cytotoxic, immune, or targeted therapies in the entire cohort, EQD2 was associated with CH-PD (p = 0.012x10−3). Evaluating EQD2 by irradiated anatomic site, total pelvic dose by EQD2 in 10 Gy increments remained significantly associated with CH-PD (OR = 1.07, p = 0.0046), as was head and neck EQD2 (OR = 1.046, p = 0.032). Conclusions: CH-PD was associated with higher radiation dose for pelvic or head and neck RT, but not other anatomic sites after controlling for systemic therapies. RT modality was not associated with CH-PD. Ongoing work will directly evaluate the bone marrow dosimetry of various treatment approaches using phantom-based modeling.


2013 ◽  
Vol 59 (2) ◽  
pp. 381-391 ◽  
Author(s):  
Shoaib Afzal ◽  
Stig E Bojesen ◽  
Børge G Nordestgaard

BACKGROUND Vitamin D deficiency has been implicated in decreased insulin secretion and increased insulin resistance, hallmarks of type 2 diabetes mellitus. We tested the hypothesis that low plasma 25-hydroxyvitamin D [25(OH)D] is associated with increased risk of type 2 diabetes in the general population. METHODS We measured 25(OH)D in 9841 participants from the general population, of whom 810 developed type 2 diabetes during 29 years of follow-up. Analyses were adjusted for sex, age, smoking status, body mass index, income, physical activity, HDL cholesterol, and calendar month of blood draw. RESULTS Lower 25(OH)D concentrations, by clinical categories or seasonally adjusted quartiles, were associated with higher cumulative incidence of type 2 diabetes (trend, P = 2×10−7 and P = 4×10−10). Multivariable adjusted hazard ratios of type 2 diabetes were 1.22 (95% CI 0.85–1.74) for 25(OH)D &lt;5 vs ≥20 μg/L and 1.35 (1.09–1.66) for lowest vs highest quartile. Also, the multivariable adjusted hazard ratio of type 2 diabetes for a 50% lower concentration of 25(OH)D was 1.12 (1.03–1.21); the corresponding hazard ratio for those ≤58 years old was 1.26 (1.15–1.41). Finally, in a metaanalysis of 16 studies, the odds ratio for type 2 diabetes was 1.50 (1.33–1.70) for the bottom vs top quartile of 25(OH)D. CONCLUSIONS We observed an association of low plasma 25(OH)D with increased risk of type 2 diabetes. This finding was substantiated in a metaanalysis.


2005 ◽  
Vol 12 (10) ◽  
pp. 1157-1163 ◽  
Author(s):  
Esmeralda Vizzi ◽  
Eva Calviño ◽  
Rosabel González ◽  
Irene Pérez-Schael ◽  
Max Ciarlet ◽  
...  

ABSTRACT The immune response elicited by the rotavirus nonstructural protein NSP4 and its potential role in protection against rotavirus disease are not well understood. We investigated the serological response to NSP4 and its correlation with disease protection in sera from 110 children suffering acute diarrhea, associated or not with rotavirus, and from 26 children who were recipients of the rhesus rotavirus tetravalent (RRV-TV) vaccine. We used, as antigens in an enzyme-linked immunosorbent assay (ELISA), affinity-purified recombinant NSP4 (residues 85 to 175) from strains SA11, Wa, and RRV (genotypes A, B, and C, respectively) fused to glutathione S-transferase. Seroconversion to NSP4 was observed in 54% (42/78) of the children who suffered from natural rotavirus infection and in 8% (2/26) of the RRV-TV vaccine recipients. Our findings indicate that NSP4 evokes significantly (P < 0.05) higher seroconversion rates after natural infection than after RRV-TV vaccination. The serum antibody levels to NSP4 were modest (titers of ≤200) in most of the infected and vaccinated children. A heterotypic NSP4 response was detected in 48% of the naturally rotavirus-infected children with a detectable response to NSP4. Following natural infection or RRV-TV vaccination, NSP4 was significantly less immunogenic than the VP6 protein when these responses were independently measured by ELISA. A significant (P < 0.05) proportion of children who did not develop diarrhea associated with rotavirus had antibodies to NSP4 in acute-phase serum, suggesting that serum antibodies against NSP4 might correlate with protection from rotavirus diarrhea. In addition, previous exposures to rotavirus did not affect the NSP4 seroconversion rate.


1978 ◽  
Vol 8 (4) ◽  
pp. 419-423
Author(s):  
P O Leinikki ◽  
I Shekarchi ◽  
P Dorsett ◽  
J L Sever

A "microgram assay" is described in which solid-phase enzyme-linked immunosorbent assay is used for the determination of specific rubella immunoglobulin G (IgG) antibody levels in micrograms per milliliter of serum. The quantitation was based on a standard curve obtained by using a reference serum, for which the specific IgG content was assayed by immunochemical purification. IgG was first purified and specific rubella antibodies were separated by an immunoadsorbent prepared by linking rubella virus antigens to Sepharose 4B. By using IgG-specific conjugate, the levels of specific rubella IgG antibodies could then be determined from clinical samples. Seronegative samples showed antibody levels less than 1 microgram/ml, whereas levels up to several hundred micrograms per milliliter were detected in some postinfection sera. The correlation between microgram antibody levels and hemagglutination inhibition titers was linear. The method offers a simple and sensitive antibody assay which could be used both for the laboratory diagnosis of acute rubella and for the evaluation of immunity.


Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4552-4552
Author(s):  
Dana E. Rollison ◽  
Neal A. Halsey ◽  
Keerti V. Shah ◽  
Raphael P. Vascidi ◽  
Kathy J. Helzlsouer

Abstract Viral infections such as HIV, EBV, and HCV have been associated with increased risk of non-Hodgkin’s lymphoma (NHL). We conducted a nested case-control study to investigate the association between prediagnostic serum antibodies to JC virus (JCV) and BK virus (BKV) and subsequent risk of NHL. Methods: Two research serum banks were established in Washington County, MD, with more than 45,000 volunteers contributing blood samples collected in 1974 and 1989. Incident cases of NHL diagnosed through 2002 (n=170) were identified among participants by linkage to population-based cancer registries. Two controls were matched to each case (n=340) on age, sex, and date of blood draw. Pre-diagnostic circulating IgG antibodies to JCV and BKV were measured using virus-like particle (VLP) enzyme-linked immunosorbant assays (ELISA). Associations between JCV and BKV antibody seropositivity and NHL were estimated using conditional logistic regression. Results: Overall, neither serum antibodies to JCV (odds ratio (OR) = 0.83, 95% confidence interval (CI) = 0.56–1.23) or BKV (OR = 0.98, 95% CI = 0.64–1.48) were associated with an increased risk of NHL. Results were similar across NHL subtypes. Examining time to diagnosis suggested a possible increase in NHL risk associated with JCV antibodies, but not BKV antibodies, among individuals diagnosed 12–20 years after the time of blood draw. Among a subset of individuals who donated blood in both 1974 and 1989, those who experienced an increase in JCV antibody levels over time were more than four times as likely to develop NHL as compared to those whose antibody levels steeply declined (OR = 4.59, 95% CI = 1.30–16.25). Conclusion: The finding of increased risk with increasing JCV antibody titers suggests a possible association between reactivation of JCV infection and subsequent NHL.


1994 ◽  
Vol 72 (04) ◽  
pp. 578-581 ◽  
Author(s):  
T McNally ◽  
S E Cotterell ◽  
I J Mackie ◽  
D A Isenberg ◽  
S J Machin

Summaryβ2 glycoprotein-I (β2GPI), a cofactor for antiphospholipid antibody (aPA) binding, binds to many anionic macromolecules including heparin. The nature of this interaction with heparin is not well understood and its effect on the purported biological functions of β2GPI is unknown.We have examined the interactions of dermatan sulphate (DS) and different pharmaceutical preparations of heparin with β2GPI by crossed immunoelectrophoresis (CIE) and investigated the effect of these agents on plasma levels of p2GPI antigen (β2GPI: Ag) by a standardised enzyme linked immunosorbent assay (ELISA). P2GPI aPA cofactor activity (β2GPI:Cof) was also measured using a modified solid phase an-ti-phosphatidylserine (aPS) ELISA. CIE results confirmed a heparin-β2GPI interaction with unfractionated (UF) heparin. β2GPI:Ag levels were unaffected by any of the preparations investigated. There were no significant differences in β2GPI:Cof activities of the samples containing LMW heparins or DS but levels of β2GPI:Cof were increased in samples containing UF sodium and calcium heparin preparations (0.5 IU/ml Monoparin, p <0.05, and 10 IU/ml Liquemin and Calcipa-rine, p <0.05).


1986 ◽  
Vol 56 (03) ◽  
pp. 250-255 ◽  
Author(s):  
C Boyer ◽  
M Wolf ◽  
C Rothschild ◽  
M Migaud ◽  
J Amiral ◽  
...  

SummaryA new solid phase enzyme-linked immunosorbent assay (ELISA) was developed for the quantitation of human Factor VII antigen (F VII Ag), using a monospecific rabbit anti-F VII antiserum. Anti-F VII F(ab′)2 fragments were adsorbed to polystyrene plates. The binding of serial dilutions of control or test plasma, containing F VII, was detected by incubation with peroxidase-labeled anti- FV II IgG followed by the addition of hydrogen peroxyde and O-phenylenediamine. This ELISA is specific, sensitive (detection limit: 0.05%) and accurate (coefficient of variation: 1.5-4% for within- and 1.6-9% for between-assays). F VII coagulant activity (F VII C) and F VII Ag were determined in large populations of controls and patients. In normal plasma (n = 38), F VII Ag ranged from 83 to 117% and the correlation coefficient between F VII Ag and F VII C was 0.94. In patients with severe (F VII C inf. 1%) congenital F VII deficiency (n = 5), F VII Ag was undetectable in two cases (inf. 0.05%) and markedly reduced (0.35 to 5.6%) in the three other cases. In patients with liver cirrhosis (n = 15), F VII Ag ranged from 21 to 59% and was in good correlation with F VII C (r = 0.84). In dicoumarol treated patients (n = 15), the levels of F VII Ag ranged from 51% to 79% and a poor correlation (r = 0.52) with F VIIC was observed. In “compensated” DIC (n = 5), levels of F VII Ag varied from 60 to 186%, with significantly higher F VII C levels (from 143 to 189%). In contrast, in “decompensated” DIC (n = 7), low F VII Ag and F VII C levels were observed (from 7 to 27%). In patients with deep-vein thrombosis (n = 25), high levels of F VII Ag (from 102 to 136%) and F VII C (from 110 to 150%) were demonstrated. In surgical patients, no significant difference was observed before and one day after intervention.


1985 ◽  
Vol 17 (10) ◽  
pp. 39-41 ◽  
Author(s):  
A. Schnattinger

Ten litres of tapwater were seeded with 200 µl (8×108 HAV particles) of a commercial (Organon Teknika) suspension of hepatitis A virus. Following WALTER and RÜDIGER (1981), the contaminated tapwater was treated with a two-stage technique for concentration of viruses from solutions with low virus titers. The two-stage technique consists of aluminium hydroxideflocculation (200 mg/l Al2(SO4)3. 18 H2O, pH 5,4-5,6) as first stage, the second stage of a lysis of aluminium hydroxidegel with citric acid/sodium citrate-buffer (pH 4,7; 1 ml/l sample), separation of viruses from the lysate by ultracentrifugation and suspension in 1 ml phosphate buffer solution (pH 7,2). A commercial solid phase enzyme-linked immunosorbent assay (ELISA) was used for the detection of HAV. HAV was detecterl in the 10.000:1 concentrates, but not in the seeded 101 samples. Approximately 4×108 of the inoculated 8×108 HAV particles were found in the 1 ml concentrates. The efficiency of detection is about 50%, the virus concentration 5000-fold. Although the percentage loss of HAV in comparison with concentration by means of membrane filtration is similar, the ultracentrifugation method yields a larger sample/concentrate ratio, so that smaller amounts of HAV can be detected more efficiently because of the smaller end-volume.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
C. J. MacDonald ◽  
A. L. Madika ◽  
G. Severi ◽  
A. Fournier ◽  
M. C. Boutron-Ruault

AbstractDyslipidaemia is a major risk factor for cardio-vascular disease, as it promotes atherosclerosis. While cross-sectional studies have identified higher serum cholesterol amongst individuals with the A blood group, there is less evidence from prospective studies whether this translates into a higher risk of dyslipidaemia that requires treatment, nor if this genetic factor interacts with smoking status. This study aimed to prospectively determine potential associations between smoking, ABO blood groups, and risk of incident dyslipidaemia requiring treatment, and to assess associations over strata of blood ABO group. We assessed associations between blood ABO group, smoking and dyslipidaemia in 74,206 women participating in the E3N cohort. We included women who did not have cardiovascular disease at baseline. Logistic regression was used to determine associations between ABO group, smoking and prevalent dyslipidaemia at baseline. Cox proportional hazard models were then used to determine if blood ABO group and smoking were associated with the risk of incident dyslipidaemia, amongst women free of dyslipidaemia at baseline. At baseline 28,281 women with prevalent dyslipidaemia were identified. Compared to the O-blood group, the non-O blood group was associated higher odds of with prevalent dyslipidaemia (ORnon-O = 1.09 [1.06: 1.13]). Amongst the women free of dyslipidaemia at baseline, 6041 incident cases of treated dyslipidaemia were identified during 454,951 person-years of follow-up. The non-O blood groups were associated with an increased risk of dyslipidaemia when compared to the O-group (HRnon-O = 1.16 [1.11: 1.22]), specifically the A blood-group (HRA = 1.18 [1.12: 1.25]). Current smokers were associated with an increased risk of incident dyslipidaemia (HR smokers = 1.27 [1.16: 1.37]), compared to never-smokers. No evidence for effect modification between smoking and ABO blood group was observed (p-effect modification = 0.45), although the highest risk was observed among AB blood group women who smoked (HR = 1.76 [1.22: 2.55]). In conclusion, the non-O blood groups, specifically the A group were associated with an increased risk of dyslipidaemia. Current smokers were associated with a 30% increased risk of dyslipidaemia. These results could aid in personalised approaches to the prevention of cardiovascular risk-factors.


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