scholarly journals Evaluation of Rapid Neutralizing Antibody Detection Test against Rabies Virus in Human Sera

2015 ◽  
Vol 43 (2) ◽  
pp. 111-116 ◽  
Author(s):  
Kieu Anh Thi Nguyen ◽  
Thu Tuyet Nguyen ◽  
Dong Vinh Nguyen ◽  
Giang Chau Ngo ◽  
Cam Nhat Nguyen ◽  
...  
2020 ◽  
Vol 12 (1) ◽  
pp. e2020016
Author(s):  
Hui Liu ◽  
Lorraine Gemmell ◽  
Rui Lin ◽  
Fengrong Zuo ◽  
Henry H. Balfour ◽  
...  

No licensed vaccine is available for prevention of EBV-associated diseases, and robust, sensitive, and high-throughput bioanalytical assays are needed to evaluate immunogenicity of gp350 subunit-based candidate EBV vaccines. Here we have developed and improved analytical tools for such a vaccine’s pre-clinical and clinical validation including a gp350-specific antibody detection assay and an EBV-GFP based neutralization assay for measuring EBV specific antibodies in human donors. The sensitivity of our previously published high-throughput EBV-GFP fluorescent focus (FFA)-based neutralization assay was further improved when guinea pig complement was supplemented using a panel of healthy human sera. Anti-gp350 antibody titers, which were evaluated using an anti-gp350 IgG ELISA assay optimized for capture and detection conditions, were moderately correlated to the FFA-based neutralization titers. Overall, these sensitive, and high-throughput bioanalytical assays are capable of characterizing the serologic response to natural EBV infection, with applications in evaluating EBV antibody status in epidemiologic studies and immunogenicity of candidate gp350-subunit EBV vaccines in clinical studies.


PEDIATRICS ◽  
1949 ◽  
Vol 4 (4) ◽  
pp. 443-453
Author(s):  
ALBERT B. SABIN

An antigen prepared from toxoplasma-infected chorioallantoic membranes, improved by centrifugation at 13,000 r.p.m. for one hour which removed a nonspecific component capable of fixing complement with certain normal sera, proved to be the preparation of choice for toxoplasmic CF tests. Though it was possible to prepare an antigen of similar potency from mouse brain, the yield from one egg is at least 10 times more than from one mouse. The peritoneal exudate of mice freed of Toxoplasma and cells contains eight times as much antigen per unit volume as a 20% extract of mouse brain or chorioallantoic membranes, but its anticomplementary properties, which cannot be removed by high speed centrifugation, can be eliminated only by extensive dilution. The highly centrifuged and diluted peritoneal exudate, however, still contains a component which fixes complement with certain normal human sera. Sixteen children, aged 5 weeks to 11 years, with clinically recognized congenital toxoplasmosis and 17 mothers, with inapparent or clinically unrecognized infection except for having given birth to children with proved toxoplasmosis, were investigated for the presence and persistence of CF antibodies. With the improved antigen and the standardized method that was used, even titers of 1:2 and 1:4 were significant and invariably associated with toxoplasmic neutralizing antibody. The sera of all 17 mothers gave positive CF tests with titers ranging from 1:4 to 1:64, the higher titers predominating during the first two years after the delivery of the infected child. The sera of 14 of the 16 children (87.5%) with clinical evidence of congenital toxoplasmosis, confirmed by neutralization tests, gave positive CF tests with titers ranging from 1:2 to 1:128, the higher titers again predominating during the first two years after birth. One of the negative results, in a 5 week old child with active infection who died three days after the serum was obtained, was associated with a high (1:4,096) dye test titer for neutralizing antibody in its own serum, and a similar dye test titer (1:4,096) and a CF titer of 1:32 in the serum of its mother. The other negative result, in a 7 year old child with a dye test titer of 1:16, is believed to represent an instance of disappearance of CF antibody after an interval of seven years. Not one serum obtained from 24 children with congenital ocular or neurologic disturbances or both, not due to toxoplasmosis, gave a positive CF test with the toxoplasmic antigen; however, among 20 of their mothers, there were four sera with titers of 1:2, 1:2, 1:8 and 1:16. Toxoplasmic CF antibody can persist for at least six years, and in some instances even longer, in individuals with clinically recognized as well as inapparent or clinically unrecognized infections. Since at least seven of the mothers, who gave birth to children with congenital toxoplasmosis, subsequently gave birth to normal children at a time when the CF antibody titers in some of them were still high (32, 32, 32, 16, 8, 4, 4), it is clear that this antibody can persist in individuals in whom the infection has been eradicated or suppressed sufficiently to prevent its congenital transmission.


2021 ◽  
Vol 12 ◽  
Author(s):  
Ilaria Varotto-Boccazzi ◽  
Alessandro Manenti ◽  
Francesca Dapporto ◽  
Louise J. Gourlay ◽  
Beatrice Bisaglia ◽  
...  

To detect and prevent emerging epidemics, discovery platforms are urgently needed, for the rapid development of diagnostic assays. Molecular diagnostic tests for COVID-19 were developed shortly after the isolation of SARS-CoV-2. However, serological tests based on antiviral antibody detection, revealing previous exposure to the virus, required longer testing phases, due to the need to obtain correctly folded and glycosylated antigens. The delay between the identification of a new virus and the development of reliable serodiagnostic tools limits our readiness to tackle future epidemics. We suggest that the protozoan Leishmania tarentolae can be used as an easy-to-handle microfactory for the rapid production of viral antigens to face emerging epidemics. We engineered L. tarentolae to express the SARS-CoV-2 receptor-binding domain (RBD) and we recorded the ability of the purified RBD antigen to detect SARS-CoV-2 infection in human sera, with a sensitivity and reproducibility comparable to that of a reference antigen produced in human cells. This is the first application of an antigen produced in L. tarentolae for the serodiagnosis of a Coronaviridae infection. On the basis of our results, we propose L. tarentolae as an effective system for viral antigen production, even in countries that lack high-technology cell factories.


1998 ◽  
Vol 36 (2) ◽  
pp. 475-480 ◽  
Author(s):  
Wolfgang Meschede ◽  
Klaus Zumbach ◽  
Joris Braspenning ◽  
Martin Scheffner ◽  
Luis Benitez-Bribiesca ◽  
...  

Cervical cancer is the most prevalent tumor in developing countries and the second most frequent cancer among females worldwide. Specific human papillomaviruses (HPVs) and, most notably, HPV types 16 and 18 are recognized as being causally associated with this malignancy. Antibodies against early HPV proteins E6 and E7 have been found more often in patients with tumors than in controls. Existing peptide enzyme-linked immunosorbent assays (ELISAs) for the detection of anti-E6 and anti-E7 antibodies in human sera have low levels of sensitivity and specificity and thus are not suitable for use as diagnostic tools. Based on highly purified recombinant native proteins, we developed four sandwich ELISAs for the detection of antibodies against HPV type 16 and 18 E6 and E7 proteins. We demonstrate their sensitivities and high degrees of specificity for cervical cancer. Among a total of 501 serum specimens from unselected patients with invasive cervical cancer, 52.9% reacted positively in at least one of the four assays. In contrast, among 244 serum specimens from control subjects without cervical cancer, only 2 reactive serum specimens (0.8%) were found. For 19 of 19 antibody-positive patients, the HPV type indicated by seroreactivity was identical to the HPV DNA type found in the tumor, which also indicates a high degree of specificity for antibody detection with respect to HPV type. In a direct comparison of 72 serum specimens from patients with cervical cancer, 56% of the specimens reacted in at least one of the four protein ELISAs, whereas 40% reacted in at least one of seven peptide ELISAs covering the four antigens. These assays could be of value for the detection of invasive cervical cancer in settings in which cytology-based early tumor screening is not available, for the clinical management of patients diagnosed with cervical cancer, and for the immunological monitoring of E6 and E7 vaccination trials.


1994 ◽  
Vol 113 (1) ◽  
pp. 137-141 ◽  
Author(s):  
E. H. Follmann ◽  
D. G. Ritter ◽  
M. Beller

SUMMARYThe purpose of this research was to determine whether trappers in northern Alaska acquired immunity to rabies virus from non-bite exposures while trapping and skinning arctic foxes (Alopex lagopus). In coastal Alaska recurring epizootics presumably provide trappers ample opportunity for contact with rabid animals. Serum neutralization analyses of blood samples collected from 26 individuals were conducted. All but three had negative rabies neutralizing antibody levels (< 0·05 I.U./ml). Two of these had previously received rabies vaccine but one individual who had trapped for about 47 years with an estimated harvest of over 3000 foxes and who had never received pre- or post-exposure rabies vaccination had a rabies serum neutralizing antibody concentration of 2·30 I.U./ml. This represents the first report of an unvaccinated person acquiring rabies virus antibody with a titre above the 0·5 I.U./ml level considered acceptable by the World Health Organization.


2019 ◽  
Vol 55 (2) ◽  
pp. 399 ◽  
Author(s):  
Kerri Pedersen ◽  
Amy T. Gilbert ◽  
Eric S. Wilhelm ◽  
Kathleen M. Nelson ◽  
Amy J. Davis ◽  
...  

2019 ◽  
Vol 32 ◽  
pp. 101502
Author(s):  
Cornelis A. De Pijper ◽  
Abraham Goorhuis ◽  
Martin P. Grobusch ◽  
Cornelis Stijnis

2019 ◽  
Vol 32 ◽  
pp. 101468 ◽  
Author(s):  
Lifeng Zhao ◽  
Faming Miao ◽  
Qi Chen ◽  
Ying Wang ◽  
Tongyan Zhao ◽  
...  

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