• Title/Summary/Keyword: immunological detection

Search Result 51, Processing Time 0.02 seconds

Rapid Detection of Salmonella enteritidis in Pork Samples with Impedimetric Biosensor: Effect of Electrode Spacing on Sensitivity

  • Kim, Gi-Young;Moon, Ji-Hea;Hahm, Bung-Kwon;Morgan, Mark;Bhunia, Arun;Om, Ae-Son
    • Food Science and Biotechnology
    • /
    • v.18 no.1
    • /
    • pp.89-94
    • /
    • 2009
  • Frequent outbreaks of foodborne illness have been increasing the awareness of food safety. Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Some immunological, rapid assays are developed, but these assays still require prolonged enrichment steps. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella entritidis in food sample. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using a semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on neutravidin-biotin binding on the surface of the IME to form an active sensing layer. To evaluate the effect of electrode gap on sensitivity of the sensor, 3 types of sensors with different electrode gap sizes (2, 5, and $10{\mu}m$) were fabricated and tested. The impedimetric biosensor could detect $10^3\;CFU/mL$ of Salmonella in pork meat extract with an incubation time of 5 min. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.

Development of a Novel Immunochromatographic Assay for Rapid Detection of VanA Ligase-Producing Vancomycin-Resistant Enterococci

  • Ji, Gil Yong;Song, Hyung Geun;Son, Bo Ra;Hong, Seung Bok;Kim, Jong Wan;Shin, Kyeong Seob
    • Journal of Microbiology and Biotechnology
    • /
    • v.24 no.3
    • /
    • pp.427-430
    • /
    • 2014
  • We developed a novel immunochromatographic assay (ICA) (EZ-Step VanA rapid kit; Dinona, Korea) for the detection of VanA ligase from vancomycin-resistant enterococci (VRE). Of eight monoclonal antibodies screened by ELISAs, the VanA ligase ICA constructed with 1H9 plus 3G11 showed the greatest reactivity. The detection limit of the kit was $6.3{\times}10^6$ CFU per test. Of 127 vancomycin-resistant microorganisms, 100 vanA VRE were positive in the VanA ligase ICA, and 27 non-vanA vancomycin-resistant isolates were negative. These results were consistent with those of the PCR analyses. Thus, our ICA is a reliable and easy-to-use immunological assay for detecting VanA-producing VRE in clinical laboratories.

A comparison of RPLA and PCR for detection of enterotoxins in methicillin-resistant Staphylococcus aureus(MRSA) strains isolated in dogs

  • Park, Son-il;Han, Hong-ryul
    • Korean Journal of Veterinary Research
    • /
    • v.39 no.4
    • /
    • pp.806-810
    • /
    • 1999
  • A multiplex-polymerase chain reaction (PCR) assay was used to detect staphylococcal enterotoxin production by 12 strains of Staphylococcus aureus isolated from clinical specimens. To evaluate the efficacy and/or sensitivity of this method, the results were compared to those obtained with the reversed passive latex agglutination kit (SET-RPLA, Denka Seiken, Japan). Of 10 strains positive by PCR were positive by RPLA but two strains, representing high sensitivity of the former method. Enterotoxin B was the most prevalent by the two methods. The kappa index between the two methods was 0.826, indicating a higher agreement and fully reliable for use. These results would suggest that sensitive, inexpensive, and relatively rapid multiplex-PCR technique may be an effective means for the detection of staphylococcal enterotoxin genes as an alternative to traditional methods such as kits or immunological methods, which depend upon the amount of enterotoxin produced.

  • PDF

Application of Immunoassay for the Detection of Gamma-Irradiated Shrimp (감마선 조사된 새우의 검지를 위한 면역분석법의 적용)

  • 이주운;육홍선;조경환;차보숙;변명우
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.30 no.4
    • /
    • pp.600-604
    • /
    • 2001
  • Immunoassay was used to study the detection method of irradiated shrimp. Sandwich ELISA was formatted with monoclonal antibody (Ab) (M-IgG) and polyclonal Abs (P-IgG) and polyclonal Abs (P-IgG) individually produced against brown shrimp tropomyosin (TPM) as an antigen. When M-IgG was used as a coating Ab to capture TPM, and P-IgG were used as reaction Ab against captured TPM could be detected in the range of 12.5 to 50 $\mu\textrm{g}$/mL. Detected concentrations of TPM from irradiated shrimp decreased dose-dependently, and the concentration of Ag by combination of irradiation with heating or freezing treatments also decreased. This results suggests the possibility for Sandwich ELISA, one of immunological analyses, to be applied for detecting irradiated shrimp.

  • PDF

Development of a Novel Immunochromatographic Assay for Rapid Detection of OXA-23 β-lactamase-producing Acinetobacter baumannii

  • Ji, Gil Young;Song, Hyung Geun;Jo, Mi Young;Hong, Seung Bok;Shin, Kyeong Seob
    • Biomedical Science Letters
    • /
    • v.22 no.2
    • /
    • pp.29-36
    • /
    • 2016
  • Among the several agents causing carbapenem resistance of Acinetobacter baumannii, the most common cause is OXA-23 ${\beta}$-lactamase, which is known to hydrolyze carbapenem. To effectively control dissemination of carbapenem-resistant Acinetobacter baumannii (CRAB), development of both rapid and easy-to-use detection methods are required. The aim of this study is to develop a novel immunochromatographic assay (ICA) for rapid detection of OXA-23 ${\beta}$-lactamase. Of the seven monoclonal antibodies (mAbs) screened by ELISA, four mAbs (4G6, 4H6, 6G4, 9A4) exhibited high reactivity. Of these four specific antibodies, the combination of 6G4/4G6 showed the greatest reactivity and this combination of mAbs (6G4/4G6 mAbs) was used to develop the OXA-23 ${\beta}$-lactamase ICA. Of 102 A. baumannii isolates tested, the OXA-23 ${\beta}$-lactamase ICA results were consistent with PCR analysis except one false positive and one false negative isolate. The overall sensitivity and specificity were 98.36% and 97.56%, respectively. In conclusion, to the best of our knowledge, we have developed the first specific antibody set to detect OXA-23 ${\beta}$-lactamase using an ICA kit. This novel ICA can be used as a reliable and easy-to-use immunological assay for detection of OXA-23 ${\beta}$-lactamase producing CRAB in clinical laboratories.

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
    • /
    • v.31 no.1
    • /
    • pp.90-96
    • /
    • 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.

Direct Detection of Escherichia coli, Staphylococcus aureus, and Salmonella spp. in Animal-derived Foods Using a Magnetic Bead-based Immunoassay

  • Kim, Jong-Hui;Yoo, Jae Gyu;Ham, Jun-Sang;Oh, Mi-Hwa
    • Food Science of Animal Resources
    • /
    • v.38 no.4
    • /
    • pp.727-736
    • /
    • 2018
  • In this study, an immuno-magnetic bead (IMB)-based assay was developed to simultaneously detect Escherichia coli, Staphylococcus aureus, and Salmonella spp. and was tested in four animal-derived foods: beef, ham, egg, and ricotta cheese. The IMB-based assay exhibited good specificity by binding to five E. coli serotypes [capture efficiency (CE) average (avg.) 90.4%], five S. aureus strains (CE avg. 91.4%), and five Salmonella serotypes (CE avg. 95.4%) but not binding to non-target bacteria (CE<10%). Furthermore, the assay detected all three pathogens with a detection limit of 10 CFU/g without the need for enrichment or additional platforms. Since the results demonstrated that the IMB-based assay can effectively separate and enrich target bacteria from a variety of animal-derived food matrixes, the assay exhibits good specificity for potential use in providing rapid, immunological, presumptive identification of pathogenic bacteria.

Respiratory Review of 2014: Tuberculosis and Nontuberculous Mycobacterial Pulmonary Disease

  • Park, Cheol Kyu;Kwon, Yong Soo
    • Tuberculosis and Respiratory Diseases
    • /
    • v.77 no.4
    • /
    • pp.161-166
    • /
    • 2014
  • Since tuberculosis (TB) remains a major global health concern and the incidence of multi-drug resistant (MDR)-TB is increasing globally, new modalities for the detection of TB and drug resistant TB are needed to improve TB control. The Xpert MTB/RIF test can be a valuable new tool for early detection of TB and rifampicin resistance, with a high sensitivity and specificity. Late-generation fluoroquinolones, levofloxacin, and moxifloxacin, which are the principal drugs for the treatment of MDR-TB, show equally high efficacy and safety. Systemic steroids may reduce the overall TB mortality attributable to all forms of TB across all organ systems, although inhaled corticosteroids can increase the risk of TB development. Although fixed dose combinations were expected to reduce the risk of drug resistance and increase drug compliance, a recent meta-analysis found that they might actually increase the risk of relapse and treatment failure. Regarding treatment duration, patients with cavitation and culture positivity at 2 months of TB treatment may require more than 6 months of standard treatment. New anti-TB drugs, such as linezolid, bedaquiline, and delamanid, could improve the outcomes in drug-resistant TB. Nontuberculous mycobacterial lung disease has typical clinical and immunological phenotypes. Mycobacterial genotyping may predict disease progression, and whole genome sequencing may reveal the transmission of Mycobacterium abscessus. In refractory Mycobacterium avium complex lung disease, a moxifloxacin-containing regimen was expected to improve the treatment outcome.

Kabuki syndrome: clinical and molecular characteristics

  • Cheon, Chong-Kun;Ko, Jung Min
    • Clinical and Experimental Pediatrics
    • /
    • v.58 no.9
    • /
    • pp.317-324
    • /
    • 2015
  • Kabuki syndrome (KS) is a rare syndrome characterized by multiple congenital anomalies and mental retardation. Other characteristics include a peculiar facial gestalt, short stature, skeletal and visceral abnormalities, cardiac anomalies, and immunological defects. Whole exome sequencing has uncovered the genetic basis of KS. Prior to 2013, there was no molecular genetic information about KS in Korean patients. More recently, direct Sanger sequencing and exome sequencing revealed KMT2D variants in 11 Korean patients and a KDM6A variant in one Korean patient. The high detection rate of KMT2D and KDM6A mutations (92.3%) is expected owing to the strict criteria used to establish a clinical diagnosis. Increased awareness and understanding of KS among clinicians is important for diagnosis and management of KS and for primary care of KS patients. Because mutation detection rates rely on the accuracy of the clinical diagnosis and the inclusion or exclusion of atypical cases, recognition of KS will facilitate the identification of novel mutations. A brief review of KS is provided, highlighting the clinical and genetic characteristics of patients with KS.

Effect of culture-promoting ingredients (CPI-107) on the culture of Mycobacterium tuberculosis (결핵균 배양에 대한 배양촉진물질(CPI-107)의 효과)

  • Seung Cheol Kim;Sezim Monoldorova;Bo-Young Jeon
    • Korean Journal of Veterinary Service
    • /
    • v.46 no.1
    • /
    • pp.29-34
    • /
    • 2023
  • Mycobacterium tuberculosis complex (M. tuberculosis complex) is a causative agent of contagious chronic disease in a wide range of mammalian hosts, mainly cattle, goat, pigs, wildlife, and humans. The definite diagnosis of tuberculosis is made based on culture of M. tuberculosis, but it takes a long time. In the present study, we analyzed whether the detection time of M. tuberculosis could be reduced when cultured in the medium containing the culture-promoting ingredients-107 (CPI-107) using the BacT/Alert 3D system, an automatic culture system. The time to detection (TTD) tended to decrease as the added concentration of CPI-107 increase. In the case of low numbers of M. tuberculosis, it decreased by 21.0% at 1.2 mg/mL of CPI-107 and by 15.9% in the case of high numbers of M. tuberculosis. In the culture using clinically isolated M. tuberculosis strains, the shortening of the culture time by CPI was more evident. In conclusion, the detection time of M. tuberculosis was shortened in the medium added with CPI-107, and this could be used for isolation, culture and drug susceptibility test of M. tuberculosis.