• Title/Summary/Keyword: immunological 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.

Development of Monoclonal Antibodies for Diagnosis of Plasmodium vivax

  • Linh, Nguyen Thi Phuong;Park, Hyun;Lee, Jinyoung;Liu, Dong-Xu;Seo, Ga-Eun;Sohn, Hae-Jin;Han, Jin-Hee;Han, Eun-Taek;Shin, Ho-Joon;Yeo, Seon-Ju
    • Parasites, Hosts and Diseases
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    • v.55 no.6
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    • pp.623-630
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    • 2017
  • Plasmodium lactate dehydrogenase (pLDH) is a strong target antigen for the determination of infection with Plasmodium species specifically. However, a more effective antibody is needed because of the low sensitivity of the current antibody in many immunological diagnostic assays. In this study, recombinant Plasmodium vivax LDH (PvLDH) was experimentally constructed and expressed as a native antigen to develop an effective P. vivax-specific monoclonal antibody (mAb). Two mAbs (2CF5 and 1G10) were tested using ELISA and immunofluorescence assays (IFA), as both demonstrated reactivity against pLDH antigen. Of the 2 antibodies, 2CF5 was not able to detect P. falciparum, suggesting that it might possess P. vivax-specificity. The detection limit for a pair of 2 mAbs-linked sandwich ELISA was 31.3 ng/ml of the recombinant antigen. The P. vivax-specific performance of mAbs-linked ELISA was confirmed by in vitro-cultured P. falciparum and P. vivax-infected patient blood samples. In conclusion, the 2 new antibodies possessed the potential to detect P. vivax and will be useful in immunoassay.

Beneficial Effects of Capparis Spinosa Honey on the Immune Response of Rats Infected with Toxoplasma Gundii

  • Hegazi, Ahmed Gaffer;Al Guthami, Fayez Mohammed;Al Gethami, Ahmed Faiz;El Fadaly, Hassan Ali
    • Journal of Pharmacopuncture
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    • v.20 no.2
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    • pp.112-118
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    • 2017
  • Objectives: The Toxoplasma gondii (T. gondii) is an intracellular opportunistic protozoan parasite that infects approximately one-third of the human population worldwide. Honey has long been used for treatment of many diseases in folk medicine. Honey has exhibited significant anthelmintic, nematicidal and anti-protozoal activities. This study was conducted to investigate the immunological patterns in rats infected with T. gondii who were treated orally with supplemented 15% Capparis spinosa honey (Saudi Arabia) for a period of 28 days. Methods: Immunoglobulin M, immunoglobulin G, and cytokines were detected by using enzyme-linked immunosorbent assays (ELISAs). In addition, the mortality and the morbidity rates were assessed. Results: Oral administration of Capparis spinosa honey as a natural food additive was experimentally shown to increase the antibody titer; furthermore, compared with the rats in the control group, the levels of the sera cytokines ($IFN-{\gamma}$, IL-1 and IL-6) were consistently higher at day 7 post-infection in the infected rats treated with oral supplements of Capparis spinosa honey. Conclusion: Orally administered supplements of Capparis spinosa honey increased both the antibody titer and the cytokines ($IFN-{\gamma}$, IL-1 and IL-6) levels in rats infected with T. gondii.

Alopecia areata in a spayed pug: clinical and immunohistochemical findings

  • Park, Seul-Gy;Lee, Chang-Min;Han, Jae-Ik;Kim, Seung-Gon;Kim, Ha-Jung;Park, Hee-Myung
    • Korean Journal of Veterinary Research
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    • v.55 no.4
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    • pp.267-269
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    • 2015
  • A two-year-old spayed female pug presented with symmetrical hyperpigmented alopecic lesions on her axillary and inguinal regions. There were no remarkable findings in dermatologic examinations and hormonal assays. Histological examination of biopsied tissues revealed prominent lymphocytic perifolliculitis along with shrunk hair follicles. Immunohistochemistry for CD3, CD79a, CD4, and CD8 showed a positive stain for CD4 antigen around hair bulbs, suggesting CD4 positive T lymphocyte infiltration. This case suggests the possibility that CD4 T lymphocyte-mediated inflammatory reaction could be a main mechanism in canine alopecia areata. Additional studies are warranted to investigate the immunological mechanism in canine species.

Analytical Techniques for Vancomycin - A Review

  • Avinash P. Sattur;Lee, Je-Hyuk;Song, Ki-Bang;T. Panda;Kim, Chul-Ho;Rhee, Sang-Ki;B. Gokul
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.3
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    • pp.153-158
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    • 2000
  • Vancomycin belongs to the vancomycin-risocetin family of glycopeptides, and is a subclass of linear sugar containg peptides composed of seven amino acids. Its strochemical configuration forms the basic of a peptidoglycon monomer. The glycosylated hexapeptide chainconsists of chloro-$\beta$-hydroxytyrosines, p-hytidoglycines, N-anthylleucine and aspartic acid forms a rigid molecular frame work and gives the difficulty in the analysis. Voncomycin in the serum samples is usually estimated by liquid chromatography and the bacterial sensitivity was genereally tested by the microbiological assay. The pressent review deals with the qualitative, quantutative, microbioligical and immunological assays and the comparison of the quantitative methods. Clinical implications of vancomycin have also been cited in the review.

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A Review of Detection Methods for the Plant Viruses

  • Jeong, Joo-Jin;Ju, Ho-Jong;Noh, Jaejong
    • Research in Plant Disease
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    • v.20 no.3
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    • pp.173-181
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    • 2014
  • The early and accurate detection of plant viruses is an essential component to control those. Because the globalization of trade by free trade agreement (FTA) and the rapid climate change promote the country-to-country transfer of viruses and their hosts and vectors, diagnosis of viral diseases is getting more important. Because symptoms of viral diseases are not distinct with great variety and are confused with those of abiotic stresses, symptomatic diagnosis may not be appropriate. From the last three decades, enzyme-linked immunosorbent assays (ELISAs), developed based on serological principle, have been widely used. However, ELISAs to detect plant viruses decrease due to some limitations such as availability of antibody for target virus, cost to produce antibody, requirement of large volume of sample, and time to complete ELISAs. Many advanced techniques allow overcoming demerits of ELISAs. Since the polymerase chain reaction (PCR) developed as a technique to amplify target DNA, PCR evolved to many variants with greater sensitivity than ELISAs. Many systems of plant virus detection are reviewed here, which includes immunological-based detection system, PCR techniques, and hybridization-based methods such as microarray. Some of techniques have been used in practical, while some are still under developing to get the level of confidence for actual use.

Diagnostic Techniques for SARS-CoV-2 Detection (SARS-CoV-2의 진단기술)

  • Kim, Jong-Sik;Kang, Na-Kyung;Park, Seon-Mi;Lee, Eun-Joo;Chung, Kyung Tae
    • Journal of Life Science
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    • v.30 no.8
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    • pp.731-741
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    • 2020
  • Coronavirus disease 19 (COVID-19) is caused by SARS-CoV-2 (Severe Acute Respiratory SyndromeCoronavirus 2). To date, seven coronaviruses that can infect humans were reported. Among them, infections with four coronavirus strains (HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1) resulted in mild symptoms such as common cold, whereas SARS-CoV and MERS-CoV caused severe symptoms and epidemics in 2002 and 2012, respectively. In the most recent, SARS-CoV-2 was first reported in Wuhan, China in December 2019 and became a notorious cause of the ongoing global pandemics. To diagnose, treat, and prevent COVID-19, the development of rapid and accurate diagnostic tools, specific therapeutic drugs, and safe vaccines essentially are required. In order to develop these powerful tools, it is prerequisite to understand a phenotype, a genotype, and life cycle of SARS-CoV-2. Diagnostic techniques have been developing rapidly around world and many countries take the fast track system to accelerate approval. Approved diagnostic devices are rapidly growing facing to urgent demand to identify carriers. Currently developed commercial diagnostic devices are divided into mainly two categories: molecular assay and serological & immunological assay. Molecular assays begins the reverse transcription step following polymerase chain reaction or isothermal amplification. Immunological assay targets SARS-CoV-2 antigen or anti-SARS-CoV-2 antibody of samples. In this review, we summarize the phenotype, genome structure and gene expression of SARS-CoV-2 and provide the knowledge on various diagnostic techniques for SARS-CoV-2.

A Current Research Insight into Function and Development of Adjuvants (면역보조제의 작용 및 개발)

  • Sohn, Eun-Soo;Son, EunWha;Pyo, SuhkNeung
    • IMMUNE NETWORK
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    • v.4 no.3
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    • pp.131-142
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    • 2004
  • In recent years, adjuvants have received much attention because of the development of purified subunit and synthetic vaccines which are poor immunogens and require adjuvants to evoke the immune response. Therefore, immunologic adjuvants have been developed and testing for most of this century. During the last years much progress has been made on development, isolation and chemical synthesis of alternative adjuvants such as derivatives of muramyl dipeptide, monophosphoryl lipid A, liposomes, QS-21, MF-59 and immunostimulating complexes (ISCOMS). Biodegradable polymer microspheres are being evaluated for targeting antigens on mucosal surfaces and for controlled release of vaccines with an aim to reduce the number of doses required for primary immunization. The most common adjuvants for human use today are aluminum hydroxide and aluminum phosphate. Calcium phosphate and oil emulsions have been also used in human vaccination. The biggest issue with the use of adjuvants for human vaccines is the toxicity and adverse side effects of most of the adjuvant formulations. Other problems with the development of adjuvants include restricted adjuvanticity of certain formulations to a few antigens, use of aluminum adjuvants as reference adjuvant preparations under suboptimal conditions, non-availability of reliable animal models, use of non-standard assays and biological differences between animal models and humans leading to the failure of promising formulations to show adjuvanticity in clinical trials. The availability of hundreds of different adjuvants has prompted a need for identifying rational standards for selection of adjuvant formulations based on safety and sound immunological principles for human vaccines. The aim of the present review is to put the recent findings into a broader perspective to facilitate the application of these adjuvants in general and experimental vaccinology.

Comparison of methods for proanthocyanidin extraction from pine (Pinus densiflora) needles and biological activities of the extracts

  • Kim, Nam-Young;Jang, Min-Kyung;Lee, Dong-Geun;Yu, Ki-Hwan;Jang, Hye-Ji;Kim, Mi-Hyang;Kim, Sung-Gu;Yoo, Byung-Hong;Lee, Sang-Hyeon
    • Nutrition Research and Practice
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    • v.4 no.1
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    • pp.16-22
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    • 2010
  • Flavonoids are known to be effective scavengers of free radicals. In particular, proanthocyanidins are flavonoids that possess cardiovascular protection, antioxidative activities, and immunomodulatory activities. Here, we evaluated proanthocyanidin contents in the total polyphenolic compounds of pine needle extracts prepared by hot water, ethanol, hexane, hot water-hexane (HWH), and hot water-ethanol (HWE). Analysis of each extract indicated that the ethanol extract contained the highest proanthocyanidin concentration. The HWH and hexane extracts also contained relatively high concentrations of proanthocyanidin. On the other hand, proanthocyanidin content analyses out of the total polyphenolic compounds indicated that the HWH extract contained the highest content. These results suggest that HWH extraction is a suitable method to obtain an extract with a high level of pure proanthocyanidins and a relatively high yield. The HWH extract possessed superior activity in diverse antioxidative analyses such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferrous ion chelating (FIC), and ferric-ion reducing power (FRAP) assays. In addition, upon assessing the effects of the pine needle extracts on macrophages (Raw 264.7 cell), the HWH extract exhibited the highest activity. In this study, we discerned an efficient extraction method to achieve relatively pure proanthocyanidins from pine needles and evaluated the biological functions of the resulting extract, which could potentially be used for its efficacious components in functional food products.

Development of Multiplex RT-PCR for Simultaneous Detection of Garlic Viruses and the Incidence of Garlic Viral Disease in Garlic Genetic Resources

  • Nam, Moon;Lee, Yeong-Hoon;Park, Chung Youl;Lee, Min-A;Bae, Yang-Soo;Lim, Seungmo;Lee, Joong Hwan;Moon, Jae Sun;Lee, Su-Heon
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.90-96
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    • 2015
  • Garlic generally becomes coinfected with several types of viruses belonging to the Potyvirus, Carlavirus, and Allexivirus genera. These viruses produce characteristically similar symptoms, they cannot be easily identified by electron microscopy (EM) or immunological detection methods, and they are currently widespread around the world, thereby affecting crop yields and crop quality adversely. For the early and reliable detection of garlic viruses, virus-specific sets of primers, including species-specific and genus-specific primers were designed. To effectively detect the twelve different types of garlic viruses, primer mixtures were tested and divided into two independent sets for multiplex polymerase chain reaction (PCR). The multiplex PCR assays were able to detect specific targets up to the similar dilution series with monoplex reverse transcription (RT)-PCR. Seventy-two field samples collected by the Gyeongbuk Agricultural Technology Administration were analyzed by multiplex RT-PCR. All seventy two samples were infected with at least one virus, and the coinfection rate was 78%. We conclude that the simultaneous detection system developed in this study can effectively detect and differentiate mixed viral infections in garlic.