Detection Of Specific Antibodies Research Articles

Anti-amphiphysin antibody-induced paraneoplastic-autoimmune encephalitis: case report and literature review

Anti-amphiphysin antibody-induced encephalitis is an immune-mediated inflammatory disease of the brain caused by onconeuronal antibodies. Amphiphysin antibodies are associated with various paraneoplastic neurological syndromes and tumours. The absence of a particular clinical pattern and pathognomonic features makes diagnosing paraneoplastic-autoimmune encephalitis complicated, especially when a malignancy is not initially apparent. Detection of specific antibodies in serum and cerebrospinal fluid remains the main diagnostic tool. Up to this date, no cases with epileptic seizures as one of the clinical manifestations of anti-amphiphysin encephalitis have been reported. In this article, we present a rare clinical case of a 54-year-old man diagnosed with autoimmune-paraneoplastic encephalitis caused by antibodies against amphiphysin which manifested with atypical clinical symptoms such as epileptic seizures, visual and perception impairment, paraesthesia, and elevated CSF protein levels with no detectable tumour.

Evaluation of an Algorithm for the Diagnosis and Therapy of Lyme Neuroborreliosis: A Follow-up Study

Background: Although Lyme Neuroborreliosis (LNB) is often seen in paediatric practice, diagnostic criteria for LNB in children are not clearly defined. The guidelines for LNB in adults are based on a combination of clinical picture, CSF pleocytosis and the detection of specific antibodies against Borrelia burgdorferi in CSF and serum. Diagnostic procedure takes several days, thus it isn´t useful in deciding for the need of prompt antibiotic treatment. Aim of study was a retrospective evaluation of an algorithm for the diagnosis and therapy of lyme’s disease, which is used since 2005 at the paediatric department of Innsbruck. Patients and Methods: All patients presenting with acute peripheral facial palsy from January 2006 to December 2014 were reviewed. The patients were diagnosed according to the algorithm, based on the criteria of the German Society of Neurology. The focus lay on evaluation of diagnosis and therapy according to the algorithm and whether overtreatment and underdiagnosis could therefore be avoided. Results: 120 patients were enrolled with peripheral facial palsy. 65 (54%) were handled as bell´s palsy and 55 (46 %) as B. burgdorferi s.l. infection. 19 cases were classified as confirmed LNB, 10 as probable and 26 as possible LNB. A total of 69 patients (58 %) were treated correctly according to the algorithm, 16 (13%) were over treated and 14 (11%) under treated with antibiotics. 21 (18%) could not be classified, according to the algorithm, due to the lack of CSF results. Although receiving proper treatment, 3 cases had a persistent defect after recovery. Conclusions: The algorithm is an appropriate diagnostic tool for the diagnosis and therapy of LNB, particularly with regard to the necessity of a prompt antibiotic treatment, and therefore helpful to avoid underdiagnosis and overtreatment.

PREVALENCE OF MEASLES IGG ANTIBODIES AMONG HEALTHCARE WORKERS IN BULGARIA

Background: Healthcare workers are at greater risk of acquiring measles than the general population. Transmission occurs from infected patients to staff and from infected staff to patients and colleagues. In inpatient and outpatient settings, susceptible patients suffering from other conditions, especially the elderly and severely ill patients in intensive care units are at a high risk of severe disease or death if infected with measles. The most effective preventive measure against measles is vaccination with two doses of a measles-containing vaccine. Aim: To perform a serological assessment of the presence of measles IgG antibodies among healthcare workers. Materials and Methods: The present study involves serological testing performed on samples from persons working in hospitals and Regional Health Inspectorates in the country. Screening started in connection with a reported measles outbreak in Bulgaria in 2019, and continued into early 2020. An indirect ELISA assay for the detection of specific measles IgG antibodies in serum samples was used. Results: The tested 152 healthcare workers were from 5 regions of the country, i.e. Sofia-capital (n=87), Burgas (n=6), Blagoevgrad (n=5), Dobrich (n=8) and Pazardzhik (n=46). Anti-measles IgG antibodies were detected in 85.5% of the participants (130/152). Negative results, indicating a possible lack of protective immunity against measles, were obtained primarily in samples from persons younger than 40 years of age. Conclusion: Screening to identify healthcare workers who lack presumptive evidence of protective immunity against measles should be reinforced, especially among younger healthcare workers, in order to minimize the risk of measles both among healthcare workers and among patients.

Recombinant SARS-CoV-2 Nucleocapsid Protein: Expression, Purification, and Its Biochemical Characterization and Utility in Serological Assay Development to Assess Immunological Responses to SARS-CoV-2 Infection.

The SARS-CoV-2 nucleocapsid protein (N) binds a single-stranded viral RNA genome to form a helical ribonucleoprotein complex that is packaged into virion particles. N is relatively conserved among coronaviruses and consists of the N-terminal domain (NTD) and C-terminal domain (CTD), which are flanked by three disorganized regions. N is highly immunogenic and has been widely used to develop a serological assay as a diagnostic tool for COVID-19 infection, although there is a concern that the natural propensity of N to associate with RNA might compromise the assay’s specificity. We expressed and purified from bacterial cells two recombinant forms of SARS-CoV-2 N, one from the soluble fraction of bacterial cell lysates that is strongly associated with bacterial RNAs and the other that is completely devoid of RNAs. We showed that both forms of N can be used to develop enzyme-linked immunosorbent assays (ELISAs) for the specific detection of human and mouse anti-N monoclonal antibodies (mAb) as well as feline SARS-CoV-2 seropositive serum samples, but that the RNA-free form of N exhibits a slightly higher level of sensitivity than the RNA-bound form to react to anti-N mouse mAb. Using the electrophoretic mobility shift assay (EMSA), we also showed that N preferentially binds ssRNA in a sequence-independent manner and that both NTD and CTD of N contribute to RNA-binding activity. Collectively, our study describes methods to express, purify, and biochemically characterize the SARS-CoV-2 N protein and to use it for the development of serological assays to detect SARS-CoV-2 infection.

LIODetect®TB-ST: Evaluation of novel blood test for a rapid diagnosis of active pulmonary and extra-pulmonary tuberculosis in IGRA confirmed patients

Because of the current limits of immunological tests in the diagnosis of tuberculosis there is a need to identify new and rapid tests that can be carried out on a large scale in endemic countries and useful in the identification of infected subjects, but also able to discriminate those with latent infection from subjects with active. We have taken into consideration and analysed the LIODetect®TB-ST Tuberculosis Rapid Test, a membrane test for the qualitative detection of specific IgG, IgA, and IgM antibodies against Mycobacterium tuberculosis, performed on serum, plasma, or whole blood.85 samples positive to QuantiFERON TB-GOLD PLUS test were processed using this test and the results obtained were concordant with clinical diagnosis.To our knowledge, the LIODetect®TB-ST Tuberculosis Rapid Test is the only test; that identifies active tuberculosis disease with high sensitivity and specificity and its use might be of help in the diagnosis of tuberculosis, especially in endemic countries.

West Nile Virus in the State of Ceará, Northeast Brazil

In June 2019, a horse with neurological disorder was diagnosed with West Nile virus (WNV) in Boa Viagem, a municipality in the state of Ceará, northeast Brazil. A multi-institutional task force coordinated by the Brazilian Ministry of Health was deployed to the area for case investigation. A total of 513 biological samples from 78 humans, 157 domestic animals and 278 free-ranging wild birds, as well as 853 adult mosquitoes of 22 species were tested for WNV by highly specific serological and/or molecular tests. No active circulation of WNV was detected in vertebrates or mosquitoes by molecular methods. Previous exposure to WNV was confirmed by seroconversion in domestic birds and by the detection of specific neutralizing antibodies in 44% (11/25) of equids, 20.9% (14/67) of domestic birds, 4.7% (13/278) of free-ranging wild birds, 2.6% (2/78) of humans, and 1.5% (1/65) of small ruminants. Results indicate that not only equines but also humans and different species of domestic animals and wild birds were locally exposed to WNV. The detection of neutralizing antibodies for WNV in free-ranging individuals of abundant passerine species suggests that birds commonly found in the region may have been involved as amplifying hosts in local transmission cycles of WNV.

Seroprevalence of antibodies to cytomegalovirus in pregnant women in the Apulia region (Italy).

SummaryIntroductionCytomegalovirus is ubiquitous and easily transmitted by contact. Following the first infection, the virus becomes latent and periodic reactivation could occur due to immunosuppression. If the infection is acquired in pregnancy, especially in the first trimester, the foetal consequences could be serious. The present study was conducted to assess the serological profile of pregnant women with respect to cytomegalovirus in Apulia from 2016 to 2019.MethodsSerum samples were tested by commercial ELISA kit for the detection of specific IgM and IgG antibodies against cytomegalovirus.ResultsThe data showed that most of the pregnant women (70.8%), especially those of ≥ 40 years of age (80.6%), has antibodies against cytomegalovirus, though these do not confer fully protective immunity against infection by different strains nor can prevent the re-activation of the latent one. Conversely, most of the youngest women are seronegative (44.4% in women < 25 years of age) and vulnerable during pregnancy.ConclusionsCurrently, cytomegalovirus screening for pregnant women is not mandatory in Italy. Considering that congenital cytomegalovirus is the leading non-genetic cause of sensorineural hearing loss, it would be extremely useful and cost-saving to screen women of childbearing age and women at early stage of pregnancy for cytomegalovirus infection in addition to increase awareness of cytomegalovirus infection and consequences among pregnant women, health care workers and the public.

Aspects of “antigen–antibody” interaction of chicken infectious bronchitis virus determined by surface plasmon resonance

Authors performed investigation on “antigen–antibody” interaction of chicken infectious bronchitis coronavirus (IBV) by a method based on the surface plasmon resonance (SPR). Presence of space-size effect related to a difference between antigen and antibody particle sizes has been theoretically grounded and experimentally proven. Herewith, the difference between responses of the SPR-sensor to specific and non-specific interactions is considerably less (up to 6.3 times) than the expected one (8 – 11 times). An impact of functionalization of sensor’s sensitive element surface, as well as acidity of buffer solution on the activity of antigen–antibody interaction was studied here. The difference between sensor’s responses to specific and non-specific interactions increased two-fold from 200 to 432ang sec due to this treatment. When changing the acidity of analyzed solution from pH7.3 to pH6.8, the corresponding difference between sensor’s responses increased by 6.3 times from 194 up to 1235ang.sec. Thus, an impact of space-size effect on interaction between IBV antigen and specific antibody can be considerably (almost in 3 times) decreased by reducing the acidity of used buffer solution. The results of our investigation can be successfully applied to develop new methods for detection of pathogens and specific antibodies using SPR.

Molecular detection and seroprevalence of classical swine fever virus from 2016 to 2018 in pigs of Mizoram, India

Classical swine fever (CSF) is a fatal endemic disease of pig population of North eastern India in particular and India in general. Present study revealed molecular detection of CSFV and seroprevalence of the disease in pig population of Mizoram, India during 2016–2018. Serum samples from apparently healthy, unvaccinated pigs were collected in collaboration with the State Animal Husbandry and Veterinary Department, Mizoram and a total of 594 serum samples from 7 districts were subjected to detection of CSFV specific antibodies by indirect ELISA. A total of 206 (34.68%) serum samples were positive for CSFV antibodies by ELISA. District wise, Saiha district showed highest seroprevalence of the disease followed by Kolasib and Serchhip. Apart from this, during the same time period, CSFV suspected samples received in the Department of Veterinary Microbiology consisting of 269 serum samples, 10 whole blood and 83 tissue samples obtained from 8 districts of Mizoram were subjected to detection of NS5b and E2 mRNA transcripts by nRT-PCR of which a total of 42 (11.60%) samples including serum (5.58%), tissues (27.71%) and whole blood (40%) were positive for the NS5b and E2 mRNA transcripts, specific for CSFV. District wise analysis revealed that Aizawl has the highest percentage of positive samples of CSFV followed by Saiha and Lawngtlai district.

A multiplex assay for the detection of antibodies to relevant swine pathogens in serum.

Livestock industry supports the livelihood of around 1.3 billion people in the world, with swine industry contributing with 30% of total livestock production worldwide. To maintain and guarantee this production, a pivotal point according to the OIE is addressing potential biohazards. To control them, permanent sero-surveillance is crucial to achieve more focused veterinary public health intervention and prevention strategies, to break the chains of transmission, and to enable fast responses against outbreaks. Within this context, multiplex assays are powerful tools with the potential to simplify surveillance programs, since they reduce time, labour, and variability within analysis. In the present work, we developed a multiplex bead-based assay for the detection of specific antibodies to six relevant pathogens affecting swine: ASFV, CSFV, PRRSV, SIV, TB and HEV. The most immunogenic target antigen of each pathogen was selected as the target protein to coat different microsphere regions in order to develop this multiplex assay. A total of 1544 serum samples from experimental infections as well as field samples were included in the analysis. The 6-plex assay exhibited credible diagnostic parameters with sensitivities ranging from 87.0% to 97.5% and specificities ranging from 87.9% to 100.0%, demonstrating it to be a potential high throughput tool for surveillance of infectious diseases in swine.

Seroprevalence of Yellow fever, Chikungunya, and Zika virus at a community level in the Gambella Region, South West Ethiopia.

Yellow fever (YF), Chikungunya (CHIK), and Zika(ZIK) are among re-emerging arboviral diseases of major public health concern. Despite the proximity of the Gambella Region to South Sudan where arboviral cases have been recorded repeatedly the current epidemiological situation is unclear in this part of southwest Ethiopia. Therefore, we conducted a community-based seroprevalence survey of YF virus (YFV), CHIK virus (CHIKV), and ZIK virus (ZIKV) infections in two selected districts. A cross-sectional study was conducted in two locations of the Gambella region (Lare and Itang) to investigate the seroprevalence of these viruses’ infections. Blood samples were collected from the study participants and screened for IgG antibodies specific to YFV and CHIKV infections using enzyme-linked immunosorbent assays (ELISA). For the detection of ZIKV specific IgG antibodies, Blockade-of-binding ELISA was used. Data were analyzed using the STATA version 13.1 Softwares. A total of 150 individuals (96 males and 54 females, age ranging from 18 to 65 years, mean age ± SD = 35.92 ± 10.99) participated and provided blood samples. Among the 150 samples 135, 90, and 150 were screened for YFV, CHIKV, and ZIKV, respectively. Hence, 2.9% (95% CI: 1.1–7.7%), 15.6% (95% CI: 9.3–24.8%), and 27.3% (95% CI: 20.7–35.3%) of samples tested positive for IgG antibodies to YFV, CHIKV, and ZIKV infections, respectively. Among the individual seropositive for ZIKV, YFV and CHIKV, only six, one and three had a history of residence outside the Gambella region respectively. Agro-pastoral occupation was significantly associated with a higher prevalence of IgG against CHIKV (AOR = 14.17; 95%CI: 2.30, 87.30) and residency in the Lare district (AOR = 11; 95%CI: 3.31, 39.81) was found to be significantly associated with a higher prevalence of IgG against ZIKV. Our findings revealed the occurrence of YFV, CHIKV and ZIKV infections in the study locations.

Complement Fixation Test for Specific Antibody Detection against Bovine Brucellosis in Selected Peasant Association of Guto Gida District, East Wollega Zone, Oromia, Ethiopia.

Brucellosis is one of the major zoonotic diseases yet prevalent in Ethiopia. It is contagious and has harmful effects on free animal movement and export. A cross-sectional study was conducted from November 2016 to April 2017 in the Loko and Uke peasant association of Guto Gida District. The goal of this study is to determine the seroprevalence and associated risk factors of bovine brucellosis. The questionnaire survey was conducted on 200 respondents to collect the livestock owner's perception of this disease. Eighty (80) blood samples were collected from grazing cattle above six months of age. Serum was tested by complement fixation test (CFT) for Brucella antibody detection. Pearson chi-square is used to assess the relation of independent variables such as sex, site, and age with seroprevalence in a 95% confidence interval if P value is less than 0.05, recoded as significant. Seroprevalence of bovine brucellosis in the present study is 22.5%. Bovine brucellosis prevalence recorded in Uke (28.6%) was higher than that in Loko (21.2%). Again bovine brucellosis is higher in males (30.4%) than females (19.2%). There is prevalence variation among site, sex, and age which is statistically insignificant (P value>0.05). Survey findings revealed that 52% (104/200) of respondents did not know the causes of abortion, while 48% (96/200) of them confirmed abortion in their herd and 64% (124/200) of respondents removed retained fetal membrane by themselves. Brucellosis is a contagious reproductive disease of cattle with zoonotic implications and needs to design and implement control measures aiming at preventing further spread of the disease.

SARS-COV-2 Infection and Specific Antibody Detection on Health Care Workers from a Military Hospital in Madrid, Spain.

This study aims to assess the COVID-19 seroprevalence in HCW at the Hospital Central de la Defensa Gómez Ulla (HCDGU) (Madrid). From 27 April to 10 June 2020 nasopharyngeal swab and serum samples from employees were processed in order to evaluate their seroprevalence and infective situation. Employees were classified according to their exposure to SARS-CoV-2 infection as high, moderate, and low exposure groups (level 1, level 2, and level 3, respectively). A specific real-time polymerase chain reaction (RT-PCR) was run to diagnose each patient, whereas the qualitative detection of IgG antibodies to SARS-CoV-2 was performed by means of an immunoassay. In total, 2781 HCW were screened. From this sample, 30 employees (1.1%) were infected with SARS-CoV-2 and 450 (16.2%) were positive to SARS-CoV-2-IgG antibodies. The seroprevalence was higher in the high exposure group.The seroprevalence of antibodies against SARS-CoV-2 among employees without any COVID-19 training was higher than in those who received COVID-19 training (14.5% vs 18.6%, P = 0.035). The seroprevalence in military and civilian personnel in level 1 was 18.2% and 20.0%, respectively (P = 0.4616), while in level 2 it was 6.0% and 16.0% (P = 0.0008) and in level 3 it was 16.7% and 10.2% (P = 0.0315). The results from the present study have shown that the high exposure group and HCW not receiving specific training against COVID-19 showed higher seroprevalence. Furthermore, the military employees from this hospital presented low percentage of seroprevalence.

Programmable, Multiplexed DNA Circuits Supporting Clinically Relevant, Electrochemical Antibody Detection.

Current health emergencies have highlighted the need to have rapid, sensitive, and convenient platforms for the detection of specific antibodies. In response, we report here the design of an electrochemical DNA circuit that responds quantitatively to multiple specific antibodies. The approach employs synthetic antigen-conjugated nucleic acid strands that are rationally designed to induce a strand displacement reaction and release a redox reporter-modified strand upon the recognition of a specific target antibody. The approach is sensitive (low nanomolar detection limit), specific (no signal is observed in the presence of non-targeted antibodies), and selective (the platform can be employed in complex media, including 90% serum). The programmable nature of the strand displacement circuit makes it also versatile, and we demonstrate here the detection of five different antibodies, including three of which are clinically relevant. Using different redox reporters, we also show that the antibody-responsive circuit can be multiplexed and responds to different antibodies in the same solution without crosstalk.

Fano Resonance-Based Blood Plasma Monitoring and Sensing using Plasmonic Nanomatryoshka.

The fast label-free detection of specific antibodies and their concentration in blood plasma is useful for many applications, e.g., in Covid-19 patients. The change in biophysical properties like the refractive index of blood plasma due to the production of antibodies during infection may be very helpful in estimating the level and intensity of infection and subsequent treatment based on blood plasma therapy. In this article, Fano resonance-based refractive index sensor using plasmonic nanomatryoshka is proposed for blood plasma sensing. The interaction between hybridized modes (bright and dark modes) in optimized nanomatryoshka leads to Fano resonance, which by virtue of steeper dispersion can confine the light more efficiently compared with Lorentzian resonance. We propose the excitation of Fano resonances in sub 100-nm size nanomatryoshka based on newly emerging plasmonic materials ZrN and HfN, and one of the most widely used conventional plasmonic material, Au. Fano resonance-based plasmonic sensors leads to sensitivity = 188.5 nm/RIU, 242.5 nm/RIU, and 244.9 nm/RIU for Au, ZrN, and HfN, respectively. The corresponding figure of merit (nm/RIU) is ~ 3.5 times 103, 3.1 times 103, and 2.8 times 103 for Au, ZrN, and HfN, respectively. Present theoretical analysis shows that refractive index sensors with high sensitivity and figure of merit are feasible using Fano modes of plasmonic nanomatryoshka.

Difficulties in differential diagnosis of inflammatory bowel diseases in patient with overlapping autoimmune hepatitis and primary sclerosing cholangitis

The article presents a clinical case of cross-syndrome of autoimmune hepatitis type 1 with primary sclerosing cholangitis. The main symptoms, laboratory and instrumental criteria of these diseases are discussed. Detection of specific antibodies is necessary for an accurate diagnosis. Interpretation of changes in the colon obtained by stepwise biopsy requires special attention. The described morphological picture does not allow to make an accurate diagnosis, it can be regarded as initial manifestations of Crohn's disease, microscopic or eosinophilic colitis, signs of worm-parasitic invasion. An example of a differential diagnostic search in such a situation is discussed.

Responsive Hydrogel Binding Matrix for Dual Signal Amplification in Fluorescence Affinity Biosensors and Peptide Microarrays.

A combined approach to signal enhancement in fluorescence affinity biosensors and assays is reported. It is based on the compaction of specifically captured target molecules at the sensor surface followed by optical probing with a tightly confined surface plasmon (SP) field. This concept is utilized by using a thermoresponsive hydrogel (HG) binding matrix that is prepared from a terpolymer derived from poly(N-isopropylacrylamide) (pNIPAAm) and attached to a metallic sensor surface. Epi-illumination fluorescence and SP-enhanced total internal reflection fluorescence readouts of affinity binding events are performed to spatially interrogate the fluorescent signal in the direction parallel and perpendicular to the sensor surface. The pNIPAAm-based HG binding matrix is arranged in arrays of sensing spots and employed for the specific detection of human IgG antibodies against the Epstein-Barr virus (EBV). The detection is performed in diluted human plasma or with isolated human IgG by using a set of peptide ligands mapping the epitope of the EBV nuclear antigen. Alkyne-terminated peptides were covalently coupled to the pNIPAAm-based HG carrying azide moieties. Importantly, using such low-molecular-weight ligands allowed preserving the thermoresponsive properties of the pNIPAAm-based architecture, which was not possible for amine coupling of regular antibodies that have a higher molecular weight.

A Novel Multiplexed Enzyme-Linked Immunosorbent Assay for the Detection of IgG Seroreactivity to Cytomegalovirus (CMV) UL144.

Infection with human cytomegalovirus (CMV) is common and may have grave consequences in transplant recipients and congenitally infected children. Diagnosis of CMV infection is based on detection of specific antibodies and molecular assays. The incorporation of CMV serological assays into diagnostic algorithms requires careful evaluation and interpretation. Very few serological assays measure CMV infection by a specific strain. We developed an enzyme-linked immunosorbent assay (ELISA) using CMV-encoded UL144 as the antigen. UL144 encodes three major genotypes, A, B, and C, and recombinants. The ELISA was developed with the three UL144 proteins and optimized as a multiplex assay. Sera from 55 positive and 59 negative CMV IgG, determined by the clinical microbiology laboratory, were used for evaluation and optimization. A cutoff optical density (OD) that distinguishes UL144 antibody-positive from antibody-negative sera was established. UL144 A, B, C, and combinations of these antigens were detected in sera. An assay threshold of 0.1 was established and, from a total of 303 sera, the overall sensitivity, specificity, and positive and negative predictive values of the multiplex ELISA were 86.72% (95% confidence interval [CI] 79.59% to 92.07%), 96.57% (92.69% to 98.73%), 94.40% (88.45% to 97.38%), and 91.60% (87.50% to 94.44%), respectively. The inter- and intraassay median coefficients of variation were 0.06 (interquartile range [IQR] 0.56, 0.2) and 0.171 (IQR 0.038, 0.302), respectively. No cross-reactivity was observed with HSV-positive CMV-negative sera. This ELISA gives simple and reproducible results for detection of anti-CMV UL144 IgG. It may assist in differentiating natural infection from CMV vaccines that lack UL144, and may provide an important tool for epidemiological studies of CMV strains.

Prevalence of Linda Virus Neutralizing Antibodies in the Austrian Pig Population.

A novel pestivirus species, termed Lateral-shaking Inducing Neuro-Degenerative Agent virus (LindaV), was discovered in a piglet-producing farm in Austria in 2015 related to severe congenital tremor cases. Since the initial outbreak LindaV has not been found anywhere else. In this study, we determined the seroprevalence of LindaV infections in the domestic pig population of Austria. A fluorophore labeled infectious cDNA clone of LindaV (mCherry-LindaV) was generated and used in serum virus neutralization (SVN) assays for the detection of LindaV specific neutralizing antibodies in porcine serum samples. In total, 637 sera from sows and gilts from five federal states of Austria, collected between the years 2015 and 2020, were analyzed. We identified a single serum showing a high neutralizing antibody titer, that originated from a farm (Farm S2) in the proximity of the initially affected farm. The analysis of 57 additional sera from Farm S2 revealed a wider spread of LindaV in this pig herd. Furthermore, a second LindaV strain originating from this farm could be isolated in cell culture and was further characterized at the genetic level. Possible transmission routes and virus reservoir hosts of this emerging porcine virus need to be addressed in future studies.

Novel gold nanoparticles functionalized Mo-polydopamine hollow sphere as an efficient quencher in conjugated microporous polymer electrochemiluminescent system

The construction of an electrochemiluminescence (ECL) biosensor achieving sensitive detection of cancer markers by improving the intensity of luminescent materials and the effect of quenchers is still under study. Herein, we designed an ECL sandwich immunosensor for sensitive detection of prostate specific antigen (PSA), which integrated gold nanoparticles functionalized Mo-polydopamine hollow sphere (Au@Mo-PDA, as a quencher) with platinum nanoparticles modified conjugated microporous polymer (Pt-CMP, as a luminophore) in the presence of triethanolamine (TEOA, as a coreactant). First, CMP with large specific surface area and high porosity was electropolymerized on glassy carbon electrodes (GCEs) employing carbazole-riched 2,6-di(9H-carbazol-9-yl) anthracene-9,10-dione as a precursor. Then, platinum nanoparticles were electrodeposited on CMP to construct a high initial ECL intensity system (signal on). Meanwhile, Pt-CMP with good biocompatibility could be used as a matrix to load a mass of prostate specific capture antibody (Ab1). When the Au@Mo-PDA labelled prostate specific detection antibody (Ab2) was introduced via sandwich immuno-reaction, the quantitative detection of PSA was realized by the quenching effect of Au@Mo-PDA since the quinone unit in Mo-PDA could annihilate the excited state of Pt-CMP* (signal-off). The proposed ECL system performs a good linearity to PSA concentration ranging from 10 fg mL−1 to 5 ng mL−1 with a low detection limit of 4.2 fg mL−1 and possesses high selectivity, stability and reproducibility. This work provides a versatile platform for the application of Mo-polydopamine based hollow sphere and conjugated microporous polymer in ECL biosensors.