• Title/Summary/Keyword: Rapid detection kit

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Development of Baccillus megaterium Disk Assay Kit for the Determination of Antibacterial Residues in Animal Tissues (식육중 잔류 향균물질의 검출을 위한 Bacillus megaterium 디스크 검사킷트 개발)

  • 손성완;조병훈;진남섭;이혜숙;윤순학;김재학;이재진;이영순
    • Journal of Food Hygiene and Safety
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    • v.11 no.4
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    • pp.315-321
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    • 1996
  • Various antimicrobial drug screen tests have been used in order to ensure food safety. However, the conventional screen tests, the Swab Test on Premises(STOP, USA), the Calf Antibiotic and Sulfa Test(CAST, USA) and the European Economic Community 4-plate Test(FPT, EU) are not sufficiently rapid or sensitive enough to detect low levels of sulfa drugs in meat. We developed a new screen test kit for the determination of the antimicrobial residues in meat called the Bacillus megaterium Disk Assay(BmDA). A comparison of BmDA with the older screen tests showed BmDA was as good as the older ones with several advantages. The new test kit is faster-it can be read in 4∼6 hours instead of 16∼18 hours. Moreover, BmDA can discriminate sulfa drugs from other antimicrobial drugs because p-aminobenzoic acid countacts the inhibiting action of sulfa drugs. Minimum detectable levels of sulfa drugs were significantly improved at the lever of 0.025*0.1 pp, compared with the level of 1.0 ppm in FPT. A comparison of BmDA with the older screen tests in HPLC confirmed meat samples exceeded the Korean tolerance value of 0.1 ppm showed BmDA was the most sensitive in the microbiological screen tests. As the microbiological screen tests have already known, a person familiar with simple laboratory techniques should have no difficulty in using it to detect antimicrobial residues in meat. This would be a simple, economic method of antimicrobial residues detection which might be succesfully used by many laboratories.

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Genetic Characterization of Molecular Targets in Korean Patients with Gastrointestinal Stromal Tumors

  • Park, Joonhong;Yoo, Han Mo;Sul, Hae Jung;Shin, Soyoung;Lee, Seung Woo;Kim, Jeong Goo
    • Journal of Gastric Cancer
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    • v.20 no.1
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    • pp.29-40
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    • 2020
  • Purpose: Gastrointestinal stromal tumors (GISTs) frequently harbor activating gene mutations in either KIT or platelet-derived growth factor receptor A (PDGFRA) and are highly responsive to several selective tyrosine kinase inhibitors. In this study, a targeted next-generation sequencing (NGS) assay with an Oncomine Focus Assay (OFA) panel was used for the genetic characterization of molecular targets in 30 Korean patients with GIST. Materials and Methods: Using the OFA that enables rapid and simultaneous detection of hotspots, single nucleotide variants (SNVs), insertion and deletions (Indels), copy number variants (CNVs), and gene fusions across 52 genes relevant to solid tumors, targeted NGS was performed using genomic DNA extracted from formalin-fixed and paraffin-embedded samples of 30 GISTs. Results: Forty-three hotspot/other likely pathogenic variants (33 SNVs, 8 Indels, and 2 amplifications) in 16 genes were identified in 26 of the 30 GISTs. KIT variants were most frequent (44%, 19/43), followed by 6 variants in PIK3CA, 3 in PDGFRA, 2 each in JAK1 and EGFR, and 1 each in AKT1, ALK, CCND1, CTNNB1, FGFR3, FGFR4, GNA11, GNAQ, JAK3, MET, and SMO. Based on the mutation types, majority of the variants carried missense mutations (60%, 26/43), followed by 8 frameshifts, 6 nonsense, 1 stop-loss, and 2 amplifications. Conclusions: Our study confirmed the advantage of using targeted NGS with a cancer gene panel to efficiently identify mutations associated with GISTs. These findings may provide a molecular genetic basis for developing new drugs targeting these gene mutations for GIST therapy.

A Quinoline carboxamide based Fluorescent Probe's Efficient Recognition of Aluminium Ion and its Application for Real Time Monitoring

  • Manivannan, Ramalingam;Ryu, Jiwon;Son, Young-A
    • Textile Coloration and Finishing
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    • v.32 no.4
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    • pp.185-192
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    • 2020
  • A novel binding site for metal ion made by designing molecule with tetrazolo quinoline with hydrazine carboxamide (TQC) and the designed molecule successfully synthesized. The probe works by selectively detecting Al3+ ion via both fluorimetric and colorimetric approach. The probe's effectiveness towards aluminium ion detection is highly sensitive and selective with no substantial interference with other competing ions. The added Al3+ ion to TQC fetched a rapid change of visual color to yellow from colorless, also the response of fluorescence turn-on. The fluorescence turn-on and color change visibly by the probe TQC with Al3+ ion credited to the ICT phenomenon (intramolecular charge-transfer transition). The likely interaction of the probe with aluminium ion has also been there predicted from ESI-MS spectral analysis results. The usefulness of the probe confirmed by practical utility by making a test kit to monitor Al3+ ion in water which showed a naked eye detection by notable color change.

Development of COVID-19 Neutralizing Antibody (NAb) Detection Kits Using the S1 RBD Protein of SARS-CoV-2 (코로나 바이러스 감염증-19의 재조합 S1 RBD 단백질을 이용한 COVID-19 바이러스의 중화항체 검사 키트의 개발)

  • Choi, Dong Ok;Lee, Kang Moon
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.3
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    • pp.257-265
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    • 2021
  • The COVID-19 virus is a β-genus virus that causes infection by mediating the angiotensin convertible enzyme 2 (ACE2) receptor, which is distributed in large numbers in the human respiratory tract. The disease requires effective post-management of antibody production by complete healers and vaccinators because there is no perfect remedy for the virus infection. This study aimed to develop recombinant proteins specifically responsive to neutralizing antibodies in clinical specimens and use them to develop a rapid diagnostic kit to diagnose neutralizing antibodies quickly and conveniently against the COVID-19 virus and confirm the possibility of commercialization through a performance evaluation. Rapid diagnostic kits using COVID-19 S1 RBD recombinant proteins can be applied to rapid diagnostic kits, with positive percentage agreement (PPA) and negative percentage agreement (NPA) of 100% and 98.3%, respectively, compared to the U.S. FDA-approved ELISA kits. If the performance of the rapid diagnostic kit is improved and neutralizing antibodies can be analyzed quantitatively using quantitative analysis equipment, it can be used as important data to predict immunity to the COVID-19 virus and determine additional vaccinations.

Clinical Evaluation of a Rapid Diagnostic Test Kit for Canine Parvovirus and Coronavirus (개 파보바이러스와 코로나바이러스 진단을 위한 신속진단키트의 임상적 유용성)

  • Chaeyeong MIN;Won-Shik KIM;Chom-Kyu CHONG;Yong LIM
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.1
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    • pp.45-51
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    • 2023
  • Canine parvovirus type 2 (CPV-2) and canine coronavirus (CCoV) are major pathogens that can induce gastroenteritis in dogs. They are highly contagious and have a high morbidity rate. There are no specific treatments available for them to date. Therefore, rapid and accurate diagnosis becomes essential. The rapid diagnostic test (RDT) for animals can be used widely in the field because it is fast and easy to use for diagnosis. Thus, this study aimed to clinically evaluate and confirm the clinical utility of CPV-2/CCoV RDT. The parameters evaluated included the limit of detection (LoD), cross-reactivity, interference, sensitivity, specificity, negative likelihood ratio (NLR), and kappa value. The results revealed that the LoD values for CPV-2 and CCoV were 9.7×10 50% tissue culture infectious dose (TCID50)/mL and 2.5×102 TCID50/mL, respectively. There was no cross-reactivity with nine pathogens or interference by interfering materials. The RDT showed a sensitivity of 90.0%, a specificity of 100.0%, NLR of 0.1, and a kappa value of 0.90 for diagnosing both viruses. In conclusion, CPV-2/CCoV RDT is useful as a screening test because of its high sensitivity, specificity, kappa value, and low NLR.

A Rapid and Sensitive Two-Site Sandwich Enzyme-Linked Immunosorbent Assay for Detection of ${\alpha}$-Fetoprotein in Human Serum

  • Jang, Jeong-Su;Kim, Jeong-Min;Chung, Gi-Hyun;Paik, Bo-Hyun;Kim, Hack-Joo
    • BMB Reports
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    • v.29 no.3
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    • pp.192-199
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    • 1996
  • A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

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Rapid and Specific Detection of Virulent V. vulnificus in Tidal Flat Sediments (갯벌 퇴적물내 병원성 Vibrio vulnificus의 신속하고 특이적인 검출)

  • Byun Ki-Deuk;Lee Jung-Hyun;Lee Kye-Joon;Kim Sang-Jin
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.168-176
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    • 2005
  • Vibrio vulnificus, one of the marine bacterial pathogens causing septicemia, was detected using molecular methods, namely, PCR and/or Southern hybridization, and real-time PCR. Extracted and purified total DNAs by using commercial kits were used as templates for PCR. Multiplex-PCR was conducted by employing three sets of primers for the genes, hemolysin (vvhA), phosphomannomutase (pmm), and metalloprotease (vvpE), for V vulnificus virulence. The presence of DMSO ($5\%$) and BSA ($0.1\%$) in PCR reaction mixture improved a detection efficiency by higher PCR band intensities. TaqMan real-time PCR was carried out by using gene segment of vvhA as a target. Detection limit of PCR/Southern hybridization without enrichments was to be around $10^2\;cells\;g^{-1}$ of sample. However, those three methods using the enrichment at $35^{\circ}C$ in APW showed high sensitivity ($2\~10\;cells\;g^{-1}$ of sediments). Highly sensitive detection of V vulnificus by real-time PCR was achieved within $5\~6$ hr, whereas the detection by PCR/Southern hybridization required about 36 hr. Thus, it was evident that real-time PCR is the most rapid and efficient method for detecting V vulnificus in tidal flat sediments.

Infection and Rapid Detection of Perkinsus sp. In Cultured Babyneck Clam, Ruditapes philippinarum from Western Coast of Korea (서해안 양식 바지락에 발생한 Perkinsus sp. 감염증과 신속검출)

  • Choi, Dong-Lim;Kwon, Jung-No;Park, Sung-Woo
    • Journal of fish pathology
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    • v.11 no.1
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    • pp.69-76
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    • 1998
  • An apicomplexan parasite, Perkinsus sp. was observed from the cultured baby clams, Ruditapes philippinarum, collected from the coast of Kochang and Taean (South Korea), where it caused seasonal mortality of clams. Several milky-white cysts were observed on the surface of gill and visceral mass of parasitised clams. The trophozoites of parasite had eccentric nucleus and proliferated by schizogony in gill, mantle, hepatopancrease and reproductive tissues, resulting in the formation of granuloma and the intensive infiltration of hemocytes in the tissues. During incubation in FTM, trophozoites increased in size, resulting in prezoosporangia which appeared as round black spheres when colored with Lugols iodine solution. The prevalence of Perkinsus sp. in clams was Kochang, 73.1%; Taean, 94.8% (during 9-mo. survey) and showed size-dependent infection. Hemacolor kit was useful to reduce time for diagnosis of the trophozoite of Perkinsus sp. that has been responsible of massive motalities in the clam.

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Evaluation of the cost-effectiveness of ASF detection with or without the use of on-field tests in different scenarios, in Sardinia

  • Cappai, Stefano;Loi, Federica;Rolesu, Sandro;Coccollone, Annamaria;Laddomada, Alberto;Sgarangella, Francesco;Masala, Sergio;Bitti, Giuseppe;Floris, Vincenzo;Desini, Pietro
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.14.1-14.10
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    • 2020
  • African swine fever (ASF) is a highly contagious disease of domestic pigs and wild boars (WBs). Without a vaccine, early antibody and antigen detection and rapid diagnosis are crucial for the effective prevention of the disease and the employment of control measures. In Sardinia, where 3 different suid populations coexisted closely for a long time, the disease persists since 1978. The recent ASF eradication plan involves more stringent measures to combat free-ranging pigs and any kind of illegality in the pig industry. However, critical issues such as the low level of hunter cooperation with veterinary services and the time required for ASF detection in the WBs killed during the hunting season still remain. Considering the need to deliver true ASF negative carcasses as early as possible, this study focuses on the evaluation and validation of a duplex pen-side test that simultaneously detects antibodies and antigens specific to ASF virus, to improve molecular diagnosis under field conditions. The main goal was to establish the specificity of the two pen-side tests performed simultaneously and to determine their ability to detect the true ASF negative carcasses among the hunted WBs. Blood and organ samples of the WBs hunted during the 2018/2019 hunting seasons were obtained. A total of 160 animals were tested using the pen-side kit test; samples were collected for virological and serological analyses. A specificity of 98% was observed considering the official laboratory tests as gold standards. The new diagnostic techniques could facilitate faster and cost-effective control of the disease.