• Title/Summary/Keyword: Serological assays

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Comparison of Laboratory Tests Applied for Diagnosing the SARS-CoV-2 Infection (SARS-CoV-2 감염의 진단에 이용되는 검사실 테스트의 비교)

  • Lee, Chang-Gun;Lee, Dongsup
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.2
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    • pp.79-94
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    • 2022
  • Due to the highly contagious nature and severity of the respiratory diseases caused by COVID-19, economical and accurate tests are required to better monitor and prevent the spread of this contagion. As the structural and molecular properties of SARS-CoV-2 were being revealed during the early stage of the COVID-19 pandemic, many manufacturers of COVID-19 diagnostic kits actively invested in the design, development, validation, verification, and implementation of diagnostic tests. Currently, diagnostic tests for SARS-CoV-2 are the most widely used and validated techniques for rapid antigen, and immuno-serological assays for specific IgG and IgM antibody tests and molecular diagnostic tests. Molecular diagnostic assays are the gold standard for direct detection of viral RNA in individuals suspected to be infected with SARS-CoV-2. Antibody-based serological tests are indirect tests applied to determine COVID-19 prevalence in the community and identify individuals who have obtained immunity. In the future, it is necessary to explore technical problems encountered in the early stages of global or regional outbreaks of pandemics and provide future directions for better diagnostic tests. This article evaluates the commercially available and FDA-approved molecular and immunological diagnostic assays and analyzes their performance characteristics.

SARS-CoV-2 Antibody Neutralization Assay Platforms Based on Epitopes Sources: Live Virus, Pseudovirus, and Recombinant S Glycoprotein RBD

  • Endah Puji Septisetyani;Pekik Wiji Prasetyaningrum;Khairul Anam;Adi Santoso
    • IMMUNE NETWORK
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    • v.21 no.6
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    • pp.39.1-39.18
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    • 2021
  • The high virulent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus that emerged in China at the end of 2019 has generated novel coronavirus disease, coronavirus disease 2019 (COVID-19), causing a pandemic worldwide. Every country has made great efforts to struggle against SARS-CoV-2 infection, including massive vaccination, immunological patients' surveillance, and the utilization of convalescence plasma for COVID-19 therapy. These efforts are associated with the attempts to increase the titers of SARS-CoV-2 neutralizing Abs (nAbs) generated either after infection or vaccination that represent the body's immune status. As there is no standard therapy for COVID-19 yet, virus eradication will mainly depend on these nAbs contents in the body. Therefore, serological nAbs neutralization assays become a requirement for researchers and clinicians to measure nAbs titers. Different platforms have been developed to evaluate nAbs titers utilizing various epitopes sources, including neutralization assays based on the live virus, pseudovirus, and neutralization assays utilizing recombinant SARS-CoV-2 S glycoprotein receptor binding site, receptor-binding domain. As a standard neutralization assay, the plaque reduction neutralization test (PRNT) requires isolation and propagation of live pathogenic SARS-CoV-2 virus conducted in a BSL-3 containment. Hence, other surrogate neutralization assays relevant to the PRNT play important alternatives that offer better safety besides facilitating high throughput analyses. This review discusses the current neutralization assay platforms used to evaluate nAbs, their techniques, advantages, and limitations.

Horses as a Potential Reservoir of Lyme Borreliosis in Jeju-do, Korea

  • Bae, Jong-Myon
    • Journal of Preventive Medicine and Public Health
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    • v.51 no.4
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    • pp.213-214
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    • 2018
  • Lyme borreliosis (LB) is the most common tick-borne infectious disease in North America, and it was designated as a national notifiable infectious disease in Korea in December 2010. While no cases in Jeju-do were recorded from 2012 to 2016, a recent survey reported that the seroprevalence of Borrelia burgdorferi using enzyme-linked immunosorbent assays in horses in Jeju-do was 19.0% (95% confidence interval, 12.0 to 28.3%). This fact suggests that horses may be a potential reservoir of LB in Jeju-do and that individuals in close contact with horses may be a high-risk group. Thus, a serological study in this high-risk group is urgently needed.

Identification and Characterization of Tobamoviruses Isolated from Commercial Pepper Seeds (시판 고추 종자에서 분리한 Tobamovirus의 동정 및 특성 조사)

  • 한정헌;손성한;나용준
    • Research in Plant Disease
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    • v.7 no.3
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    • pp.164-169
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    • 2001
  • Two Tobamoviruses showing different local lesion types on Nicotiana glutinosa was isolated from commercial pepper seeds. These viruses were designated Tobamovirus-6 (T-6) and Tobamovirus-19 (T-19). The biological and serological assays revealed that T-6 and T-19 were closely related to Pepper mild mottle virus (PMMoV) and Tomato mosaic virus (ToMV), respectively, The isolates also had low similarity in the array of viral coat protein gene sequences, of which T-19 was most identical to known strains of ToMV, while T-6 was closely related to PMMoV.

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Detection of Respiratory Viral Pathogens and Mycoplasma spp from Calves with Summer Pneumonia in Korea

  • Park, Jung-hoon;Kim, Doo
    • Journal of Veterinary Clinics
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    • v.36 no.4
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    • pp.185-189
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    • 2019
  • Respiratory pathogens of calves including bovine parainfluenza type 3 virus (BPI3V), bovine respiratory syncytial virus (BRSV), infectious bovine rhinotracheitis virus (IBRV) and Mycoplasma spp is well-known for winter pathogens. However, there are no studies about summer pneumonia pathogens of calves in Korea. The aim of this study was to detect respiratory pathogens from calves with summer pneumonia. Eighty calves from 5 regions were chosen and their nasal swabs were used to detect respiratory pathogens with real-time PCR. Mycoplasma spp was major primary respiratory pathogens in calves with summer pneumonia. Although, the detection rates of respiratory viruses were very low, serological assays showed that respiratory viruses exist widely in farms.

Isolation, identification and serological investigation of Actinobacillus pleuropneumoniae in slaughtered pigs (도축돈에서의 Actinobacillus pleuropneumoniae 분리, 동정 및 감염률 조사)

  • Kim, Kyung-Eon;Ku, Kyung-Nyer;Ko, Jae-Hyung;Moon, Hyeong-Jun;Choi, Kwon-Rag;Song, Eun-Ah;Park, Mi-Young
    • Korean Journal of Veterinary Service
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    • v.36 no.3
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    • pp.181-186
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    • 2013
  • This study was conducted to isolate the Actinobacillus pleuropneumoniae (APP) and to find out the distribution of 15 serovars mainly in southern Gyeonggi province, Korea. From July 2011 to Nov. 2012, a total of 2,204 slaughter pigs (110 herds) were inspected for evaluation of APP like pneumonic lesions. 48 (33.8%) APP strains were isolated from the 142 lungs and identified using PCR assays (cps, apx/omlA, biovar). Consequently, the serotype ratio were as in the following; type2 41.7% (n=20), type5 33.3% (n=16), type12 10.4% (n=5), type1 6.2% (n=3), type4 and 7 2.1% (n=1) and unknown 4.2% (n=2). Also serological test was implemented for 452 (83 herds) serum samples randomly collected from above slaughter pigs using commercial ELISA kits. The positive ratio of each serotype for tested pigs were 19.1% (77/404) on [2], 7.1% (32/452) on [3, 6, 8], 6.9% (28/404) on [5a, 5b], 6.2% (28/452) on [4, 7], 2.8% (9/320) on [12], 2.0% (9/452) on [1, 9, 11] and 0.0% (0/452) on [10]. And 49.3% (223/452) of pigs were positive on apxIV antibody. On the basis of latter screening test, the infected farm ratio accounted for 71.1% (59/83) and that was much higher than previously reported data.

Enterotoxigenic Escherichia coli Isolated from Diarrheal Patients (환자대변에서 장관병원성 대장균의 분리 -독소원성 대장균의 검출에 대하여-)

  • Kim, Yung-Bu;Yang, Hak-Do
    • The Journal of the Korean Society for Microbiology
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    • v.19 no.1
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    • pp.65-71
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    • 1984
  • The author attempted serological typing with the slide agglutination and the production of heat-labile enterotoxin (LT) and heat-stable enterotoxin (ST) by the enterotoxingenic eschenchia coli 446 strains isolated from diarrheal patients. The enterotoxingenic E.coli, producing LT and/or St, were detected by use of assays in the reversed passive hemagglutination(RPHA) and the suckling mouse method. The results obstained were as follows: 1. Of 446 strains isolated, 88 strains(19.73%) produced LT, while 224 strains(50.22%) produced ST, 134(30.05%) produced both LT-ST simultaneously. Serological typing were typed into 06, 08, 020, 025, 027, 0126, 0148 and 0159. Serotype 06 had the highest incidence of 26.46% followed by 0148(18.16%), 027(14.57%); 025(10.99%), 0159(4.03%), 0126(3.59%), 020(1.12%) and 08(0.45%). 2. Serotype 06, 08 and 025 almost always produced both LT and ST, whereas serotypes 020, 027 and 0148 almost always produced only ST. And serotypes 06, 025, 0126 and 0159 almost always produced LT or ST. 3. Of 134 LT-ST positive strains, 115 strains were serotype 06, 3 strains were 025, 2 strains were 08, and 14 strains were 0 untypable. 4. Of 224 ST positive strains, 65 strains were serotype 027, 81 strains were 0148, 16 strains were 0159, and 42 strains were 0 untypable. 5. Of 88 LT positive strains, 45 strains were serotype 025, 5 strains were 0126, 2 strains were 0159, and 42 strains were 0 untypable.

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Molecular Method Confirms Canine Leishmania Infection Detected by Serological Methods in Non-Endemic Area of Brazil

  • Riboldi, Emeline;Carvalho, Flavio;Romao, Pedro Roosevelt Torres;Barcellos, Regina Bones;Bello, Graziele Lima;Ramos, Raquel Rocha;de Oliveira, Rosemari Terezinha;Araujo, Joao Pessoa Junior;Rossetti, Maria Lucia;Dallegrave, Eliane
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.11-19
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    • 2018
  • In Brazil, visceral leishmaniasis (VL) is expanding and becoming urbanized, especially in non-endemic areas such as the State of Rio Grande do Sul. Considering that infected dogs are the main reservoir for zoonotic VL, this study evaluated the prevalence of canine visceral leishmaniasis (CVL) in the metropolitan area of Porto Alegre, a new area of expansion of VL in Brazil. Serum and plasma from 405 asymptomatic dogs from the municipalities of Canoas (n=107), $S\tilde{a}o$ Leopoldo (n=216), and Novo Hamburgo (n=82) were tested for CVL using immunochromatographic ($DPP^{(R)}$) and ELISA $EIE^{(R)}$ assays (2 assays officially adopted by the Brazilian government for the diagnosis of CVL) and real-time PCR to confirm the results. There was no agreement among serological and real-time PCR results, indicating that the Leishmania infection in asymptomatic animals with low parasite load, confirmed by negative parasitological tests (smears and parasite culture), need to be evaluated by molecular methods. The prevalence of LVC in the metropolitan region of Porto Alegre, confirmed by real-time PCR was 4% (5.6% in Canoas and 4.6% in $S\tilde{a}o$ Leopoldo). The use of molecular method is essential for accurate diagnosis of CVL, especially in asymptomatic dogs in non-endemic areas.

Expression of Lily mottle virus Coat Protein and Preparation of IgY Antibody against the Recombinant Coat Protein

  • Yoo, Ha Na;Jung, Yong-Tae
    • Horticultural Science & Technology
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    • v.32 no.4
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    • pp.544-549
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    • 2014
  • Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf and bulb samples showing characteristic symptoms of virus infection were collected in 2012, and 80 field samples were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The infection frequencies were 79% for LMoV, 5% for LSV, and 3% for CMV. The LMoV coat protein gene was amplified and cloned into the pET21d(+) expression vector to develop serological diagnostic tools to detect LMoV. The resulting carboxy-terminal His-tagged coat proteins were expressed in Escherichia coli strain BL21 (DE3) by induction with IPTG. The recombinant proteins were purified using Ni-NTA agarose beads and used as an antigen to produce polyclonal antibodies in laying hens. The resulting egg yolk immunoglobulin (IgY) specifically recognized LMoV from infected plant tissues in immunoblotting assays and had comparable sensitivity to that of a mammalian antibody. In addition, method of immunocapture RT-PCR using this IgY was developed for sensitive, efficient, and rapid detection of LMoV. Based on these results, large-scale bulb tests and detection of LMoV in epidemiological studies can be performed routinely using this IgY. This is the first report of production of a polyclonal IgY against a plant virus and its use for diagnosis.