• Title/Summary/Keyword: Rapid detection kit

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Development of a biosensor from aptamers for detection of the porcine reproductive and respiratory syndrome virus

  • Kuitio, Chakpetch;Rasri, Natchaya;Kiriwan, Duangnapa;Unajak, Sasimanas;Choowongkomon, Kiattawee
    • Journal of Veterinary Science
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    • v.21 no.5
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    • pp.79.1-79.12
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    • 2020
  • Background: Recently, the pork industry of Thailand faced an epidemic of highly virulent strains of porcine reproductive and respiratory syndrome virus (PRRSV), which spread throughout Southeast Asia, including the Lao People's Democratic Republic and Cambodia. Hence, the rapid and on-site screening of infected pigs on a farm is essential. Objectives: To develop the new aptamer as a biosensor for detection PRRSV which are rapid and on-site screening of infected pig. Methods: New aptamers against PRSSV were identified using the combined techniques of capillary electrophoresis, colorimetric assay by gold nanoparticles, and quartz crystal microbalance (QCM). Results: Thirty-six candidate aptamers of the PRRSV were identified from the systematic evolution of ligands by exponential enrichment (SELEX) by capillary electrophoresis. Only 8 out of 36 aptamers could bind to the PRSSV, as shown in a colorimetric assay. Of the 8 aptamers tested, only the 1F aptamer could bind specifically to the PRSSV when presented with the classical swine fever virus and a pseudo rabies virus. The QCM was used to confirm the specificity and sensitivity of the 1F aptamer with a detection limit of 1.87 × 1010 particles. Conclusions: SELEX screening of the aptamer equipped with capillary electrophoresis potentially revealed promising candidates for detecting the PRRSV. The 1F aptamer exhibited the highest specificity and selectivity against the PRRSV. These findings suggest that 1F is a promising aptamer for further developing a novel PRRSV rapid detection kit.

Comparison of chemiluminescence immunoassay (CIA) and immunochromatography assay(ICA) for Detecting HBsAg and Anti-HBs (B형 간염항원 및 항체 검사를 위한 화학발광면역검사법(CIA)과 면역크로마토그래피법(ICA)의 비교분석)

  • Kim, Hye-Jeong;Kim, Eun-Hee
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.7
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    • pp.3419-3424
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    • 2013
  • Diverse immunoassays including a chemiluminescent immunoassay (CIA) are used to detect hepatitis B surface antigen (HBsAg) and antibody (anti-HBs). Recently, an increasing number of institutions have been utilizing an immunochromatography assay (ICA), which is easy to use. In this study, We evaluated ICA kits for the rapid detection of HBsAg and anti-HBs by comparing them with a CIA. A total of 120 serum hospital samples, were collected for the whole month, were assayed using ICA kit. The Concordance rate, sensitivity, specificity, positive predictive value and negative predictive value of the ICA for HBsAg based on CIA results were 97%, 97%, 100%, 100%, and 96.8%, respectively. The diagnostic performances of the ICA for Anti-HBs were 90%, 90%, 93.3%, 93.1%, and 90.3%, respectively. The ICA kit failed to detect HBsAg and anti-HBs in low reactive samples. The ICA kits for the rapid detection of HBsAg might be recommended for interpreted with caution and dual analysis in the clinical laboratory.

Comparison of Acid-Fast Staining, PCR, LCR, PCR-Hybridization for Detection of Mycobacterum Tuberculosis in Clinical Specimens (임상 검체에서 결핵균 검출을 위한 항산성염색, PCR, LCR, PCR-Hybridization 검사법 간의 비교)

  • Choi, Jong-Rak;Lim, Jong-Baeck;Kim, Hyung-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.281-289
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    • 2000
  • Background : Mycobacterial culture is a confirmatory test to detect. M. tuberculosis, but it takes at least 6 weeks to diagnose. PCR is a rapid and sensitive method, but it is known that PCR has a high false positive rate due to contamination, and a high false negative rate due to inhibitors. It is also known that LCR and PCR-Hybridization, recently developed methods, are more specific methods than PCR in terms of detecting M. tuberculosis. In this study, we estimated the clinical utility of in house PCR, LCR and PCR-Hybridization for the detection of M. tuberculosis. Methods : We evaluated 75 specimens, upon which M. tuberculosis culture based testing was requested, by PCR LCR, and PCR-Hybridization and compared results. Mycobacterial culture was performed on 3% Ogawa media for 8 weeks, and an in house PCR, LCx Mycobacterium tuberculosis assay kit (Abbott Laboratories, North Chicago, III) and the AMPLICOR M. tuberculosis test kit (Roche Molecular Systems, Inc. Branchburg, NJ. USA). Results : In the view of the culture results, the sensitivities of the three tests were 40%, 80%, and 100% and their specificities were 98.6%, 94.3%, and 94.3%. Conclusion : LCR and PCR-Hybridization are rapid and sensitive methods for detecting M. tuberculosis in clinical laboratories.

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Trends in Rapid Detection Methods for Food-borne pathogenic Microorganisms by Using New Technologies (신기술 이용 식중독균 신속검출법 개발 동향 분석)

  • Kim, Hyun-Joo;Kim, Yong-Soo;Chung, Myung-Sub;Oh, Deog-Hwan;Chun, Hyang-Sook;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.376-387
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    • 2010
  • Recently, speedy, convenient and easy detection technologies have been developed rapidly and on the contrary, studies on development of traditional detectors applying biochemical characteristics has gradually been decreased. This review examined trend in current studies on detection of food-borne pathogenic microorganisms in the fields of selective media, immuno-assay, Polymerase Chain Reaction (PCR), microarray, terahertz spectroscopy & imagination and so on. Most traditional methods to detect the organisms from food matrix rely on selective media and such a method have disadvantages like long time requirement and distinguishing one species only from each selective medium although they are highly economical. Various new convenient methods such as Enzyme Linked Immuno-sorbent Assay (ELISA), paper-strip kit, fluoroimmunoassay etc. have been developed. The most ideal method for detecting food-borne pathogenic microorganisms in foods should be accurate, convenient, rapid and economical. Additionally, it is needed that capabilities of quantitative analysis and automation to be applied to industries.

On the study of Chemical Disaster Cause Chemical Detection Process (화학재난 현장에서의 사건원인 화학물질 탐지절차 연구)

  • Kim, Sungbum;Ahn, Seungyoung;Lee, Jinhwan
    • Journal of the Society of Disaster Information
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    • v.10 no.3
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    • pp.452-457
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    • 2014
  • The event of a Chemical disaster response personnel are causative events quickly Appearance & residual contaminant concentrations, should be identified accurately. In addition, the chemical disaster response procedure appropriate progress in the field of Chemical Composition and contaminant concentrations in order confirmation is essential. Use in the field to using the characteristics of each equipment. on-Site response equipment can not verify all the chemicals, materials detection, limited by each equipment. Detection range of equipment & specific materials should be considered complementary. In this study, using the equipment on-site detection of detection kit and detector tube, electronic detection equipment utilized for the rapid response procedure for helping a person to cope.

Validation of a Real-Time RT-PCR Method to Quantify Newcastle Disease Virus (NDV) Titer and Comparison with Other Quantifiable Methods

  • Jang, Juno;Hong, Sung-Hwan;Kim, Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.100-108
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    • 2011
  • A method for the rapid detection and quantification of Newcastle disease virus (NDV) produced in an animal cell culture-based production system was developed to enhance the speed of the NDV vaccine manufacturing process. A SYBR Green I-based real-time RT-PCR was designed with a conventional, inexpensive RT-PCR kit targeting the F gene of the NDV LaSota strain. The method developed in this study was validated for specificity, accuracy, precision, linearity, limit of detection (LOD), limit of quantification (LOQ), and robustness. The validation results satisfied the predetermined acceptance criteria. The validated method was used to quantify virus samples produced in an animal cell culture-based production system. The method was able to quantify the NDV samples from mid- or late-production phases, but not effective on samples from the early-production phase. For comparison with other quantifiable methods, immunoblotting, plaque assay, and tissue culture infectious dose 50 ($TCID_{50}$) assay were also performed with the NDV samples. The results demonstrated that the real-time RT-PCR method is suitable for the rapid quantification of virus particles produced in an animal cell-culture-based production system irrespective of viral infectivity.

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
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    • v.39 no.4
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    • pp.806-810
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    • 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.

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Comparative Assessment of Diagnostic Performances of Two Commercial Rapid Diagnostic Test Kits for Detection of Plasmodium spp. in Ugandan Patients with Malaria

  • Bahk, Young Yil;Park, Seo Hye;Lee, Woojoo;Jin, Kyoung;Ahn, Seong Kyu;Na, Byoung-Kuk;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.56 no.5
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    • pp.447-452
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    • 2018
  • Prompt diagnosis of malaria cases with rapid diagnostic tests (RDTs) has been widely adopted as an effective malaria diagnostic tool in many malaria endemic countries, primarily due to their easy operation, fast result output, and straightforward interpretation. However, there has been controversy about the diagnostic accuracy of RDTs. This study was conducted to evaluate the diagnostic performances of the 2 commercially available malaria RDT kits, RapiGEN Malaria Ag Pf/Pv (pLDH/pLDH) and Asan $EasyTest^{TM}$ Malaria Ag Pf/Pv (HRP-2/pLDH) for their abilities to detect Plasmodium species in blood samples collected from Ugandan patients with malaria. To evaluate the diagnostic performances of these 2 RDT kits, 229 blood samples were tested for malaria infection by microscopic examination and a species-specific nested polymerase chain reaction. The detection sensitivities for P. falciparum of Malaria Ag Pf/Pv (pLDH/pLDH) and Asan $EasyTest^{TM}$ Malaria Ag Pf/Pv (HRP-2/pLDH) were 87.83% and 89.57%, respectively. The specificities of the 2 RDTs were 100% for P. falciparum and mixed P. falciparum/P. vivax infections. These results suggest that the 2 RDT kits showed reasonable levels of diagnostic performances for detection of the malaria parasites from Ugandan patients. However, neither kit could effectively detect P. falciparum infections with low parasitaemia (<$500parasites/{\mu}l$).

Rapid Detection of Pathogens Associated with Dental Caries and Periodontitis by PCR Using a Modified DNA Extraction Method (PCR을 이용한 치아우식증 및 치주염 연관 병원체의 빠른 검출)

  • Kim, Jaehwan;Kim, Miah;Lee, Daewoo;Baik, Byeongju;Yang, Yeonmi;Kim, Jaegon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.41 no.4
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    • pp.292-297
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    • 2014
  • DNA extraction is a prerequisite for the identification of pathogens in clinical samples. Commercial DNA extraction kits generally involve time-consuming and laborious multi-step procedures. In the present study, our modified DNA isolation method for saliva samples allows for the quick detection of pathogens associated with dental caries or periodontitis by PCR within 1 h. To release DNA from the bacteria, 1 min of boiling was adequate, and the resulting isolated DNA can be used many times and is suitable for long term storage of at least 13 months at $4^{\circ}C$, and even longer at $-20^{\circ}C$. In conclusion, our modified DNA extraction method is simple, rapid, and cost-effective, and suitable for preparing DNA from clinical samples for PCR for the rapid detection of oral pathogens from saliva.

Development of Lateral Flow Immunoassay for Antigen Detection in Human Angiostrongylus cantonensis Infection

  • Chen, Mu-Xin;Chen, Jia-Xu;Chen, Shao-Hong;Huang, Da-Na;Ai, Lin;Zhang, Ren-Li
    • Parasites, Hosts and Diseases
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    • v.54 no.3
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    • pp.375-380
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    • 2016
  • Angiostrongyliasis is difficult to be diagnosed for the reason that no ideal method can be used. Serologic tests require specific equipment and are not always available in poverty-stricken zone and are time-consuming. A lateral flow immunoassay (LFIA) may be useful for angiostrongyliasis control. We established a LFIA for the diagnosis of angiostrongyliasis based on 2 monoclonal antibodies (mAbs) against antigens of Angiostrongylus cantonensis adults. The sensitivity and specificity were 91.1% and 100% in LFIA, while those of commercial ELISA kit was 97.8% and 86.3%, respectively. Youden index was 0.91 in LFIA and 0.84 in commercial ELISA kit. LFIA showed detection limit of 1 ng/ml of A. cantonensis ES antigens. This LFIA was simple, rapid, highly sensitive and specific, which opened an alternative approach for the diagnosis of human angiostrongyliasis.