• 농업과학연구
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• 제46권3호
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• pp.549-557
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• 2019

Staphylococcal enterotoxin is a very common cause of food poisoning. Conventional detection methods for the toxin including enzyme-linked immunosorbent assays (ELISAs), chemiluminescence (ECL), and polymerase chain reaction (PCR) assays require a lot of time, efforts, and expert technicians. Lateral flow strip kits have shown great potential for the rapid detection of foodborne pathogens. The lateral flow strip kit is widely used in clinical settings because it is easy to use, fast, and cost effective. A typical lateral flow strip kit uses colloidal gold to generate a visual signal. However, the lateral flow strip kit based on colloidal gold has limited sensitivity to fulfill food safety regulation requirements. This study was performed to develop a rapid test kit for pathogenic staphylococcal enterotoxin B (SEB) in food samples. The rapid detection kit was fabricated based on a nitrocellulose lateral-flow strip. Cellulose nano beads and SEB antibodies were used as the tag and receptor, respectively, to improve the detection performance. Manually spotted SEB antibody and anti-rabbit antibody on the surface of the nitrocellulose membrane were used as test and control spots, respectively. The feasibility of the rapid test kit to detect SEB in samples was evaluated. The sensitivity of the kit was 10 ng/mL SEB spiked in PBS. Additionally, the rapid test kit could detect 1 ng/mL of SEB in chicken meat extract.

• 식물병연구
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• 제20권1호
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• pp.15-20
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• 2014

A rapid, user-friendly and simple immune-chromatographic dipstick kit named 'rapid immune-gold strip' (RIGS) kit was developed in a novel single strip format to detect on-site detection of Tomato spotted wilt virus (TSWV). Immunoglobulin G (IgG) from polyclonal antisera raised in rabbits against TSWV was purified through protein-A affinity chromatography and then the purified TSWV-IgG was conjugated to colloidal gold nano-particles which served as a test line on nitrocellulose membrane. Protein A that non-specifically binds to TSWV antibody was used as a control line on the same strip. The diagnosis process with the TSWV-RIGS involves simply grinding the suspect plant sample in a bag that contains the extraction buffer and inserting the strip the bag. Results can be seen in 2-5 minutes. The flow of the complexes of gold particles coated with TSWV-IgG and a crude sap from TSWV-infected pepper, tobacco and tomato plants resulted in intensive color formed on the test lines proportional to the concentrations of TSWV. The RIGS-TSWV kit did not show any cross-reactions against other tomato-infecting viruses unrelated to TSWV. These results indicate that the TSWV-RIGS kit is highly sensitive and is not required for laboratory training and experience prior to testing. The TSWV-RIGS kit is suitable for on-site detection of suspect TSWV-infected plants as well as for laboratory diagnosis.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권2호
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• pp.127-131
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• 2004

A low-cost, simple strip reader system using a linear movement mechanism of CD-ROM deck has been developed to characterize a lateral flow membrane-based immunochromatographic assay. The test strip reader was assembled by a CD-ROM deck and home-made optical head especially designed for immunoassays. The optical head for detecting reflected light from the test strip surface consists of green light-emitting diode, large area silicon photodiode, and anodized aluminum mounting block providing a slit structure for cutting light from the LED. The stepping motor of the deck was operated in the full step mode, whose distance of each reading point is about 0.15mm. The performance of the strip reader was tested by analysis of HBV(hepatitis B virus) antigen test kit. This strip reader can be useful for inexpensive, disposable, and membrane-based assays that provide visual evidence of the presence of an analyte in a liquid sample.

• Journal of Biosystems Engineering
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• 제36권2호
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• pp.140-146
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• 2011

This study was performed to develop a rapid test kit for pathogenic Salmonella in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and biotin conjugated Salmonella antibodies were used as a tag and a receptor, respectively. Manually spotted Salmonella antibody and Neutravidin on nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Salmonella typhimurium in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Salmonella spiked in PBS. Also, the rapid test kit could detect $10^6$ cfu/mL of Salmonella in chicken meat extract.

• 농업과학연구
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• 제40권2호
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• pp.139-146
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• 2013

This study was performed to develop a rapid test kit for pathogenic Staphylococcus in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and Staphylococcus antibodies were used as a tag and a receptor, respectively. Manually spotted Staphylococcus antibody and anti-mouse antibody on the surface of nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Staphylococcus aureus in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Staphylococcus spiked in PBS. Also, the rapid test kit could detect $10^5$ cfu/mL of Staphylococcus in chicken meat extract.

• 한국치위생학회지
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• 제3권2호
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• pp.197-208
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• 2003

This study was implemented for 84 students of dental hygiene to show the correlation between dental caries experience and improved caries activity test. Dental caries experience for the sample groups was examined and stimulative saliva secreted for 5 minutes was collected into the tube to check saliva secretion rate. Dentocult LB test was executed to observe Lactobacilli colonies after 96 hour cultivation of culture slides moistened with stimulative saliva. Dentocult SM test(screening strip, site strip) was done to measure SM colonies distribution after 48 hour cultivation of culture strips applied with collected saliva and dental plaque respectively, and salivary buffering capacity was checked by means of Dentobuff strip kit. Following conclusions are obtained after examining the relation between Dentocult LB, Dentocult SM, Dentobuff strip test results and DMFT index, salivary secretion rate. 1. Showed no significant difference between Dentocult LB test results and DMFT index, salivary secretion rate. 2. Showed no significant difference between Dentocult SM(screening strip) test results and DMFT index, salivary secretion rate. 3. Showed significant difference between Dentocult SM(site strip) test results and DMFT index(pE0.05), but showed no significant difference between Dentocult SM(site strip) test results and salivary secretion rate. 4. Showed no significant difference between Dentobuff strip test results and DMFT index, but showed a very wide difference between Dentobuff strip test results and salivary secretion rate(pE0.01).

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1152-1161
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• 2007

An immunochromatography (ICG) strip test based on a monoclonal antibody for the rapid detection of L. monocytogenes in meat and processed-meat samples was developed in this study. A monoclonal antibody (MAb) specific to L. monocytogenes was produced from cloned hybridoma cells (FKLM-3B12-37) and used to develop an ICG strip test. The antibody showed a stronger binding to L. monocytogenes than other Listeria species, and a weak cross-reaction to S. aureus based on an ELISA. The detection limit of the ICG strip test was $10^5\;cell/ml$. In total, 116 meat and processed-meat samples were collected and analyzed using both the ICG strip test and a PCR. The ICG strip test and PCR indicated L. monocytogenes contamination in 34 and 27 meat samples, respectively. The 7 meat samples not identified as L. monocytogenes positive by the PCR were also tested using an API kit and found to be contaminated by Listeria species. In conclusion, the ICG strip test results agreed well with those obtained using the PCR and API kit. Thus, the developed ICG has potential use as a primary screening tool for L. monocytogenes in various foods and agricultural products, generating results within 20 min without complicated steps.

• 대한의생명과학회지
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• 제4권2호
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• pp.103-112
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• 1998

과산화 효소와 포도당 산화효소의 효소반응을 이용하여 혈액 중 당을 측정하는 혈당 측정 용 스트립을 개발하였다. 멤브레인에 포도당 산화효소와 과산화 효소, 색원체를 건조처리하면 혈당이 멤브레인에 처리된 포도당 산화효소와 즉각 반응하여 과산화 수소를 발생하고 발생된 과산화 수소가 과산화 효소와 반응하여 착색물을 형성한다. 제조된 스트립은 혈액과 접촉하면 반응시간 2∼3분 이내에 0∼800mg/d1의 혈당에 대하여 농도에 따라 연 녹, 청 녹, 짙은 청색을 나타내며 이때 민감도는 40 mg/dl이었다. 육안용 혈당 스트립을 이용하여 정상을 포함한 당뇨 환자의 혈당 농도를 측정한 결과 Ames사와 BM사에서 시판하고 있는 제품과 유사한 결과를 얻을 수 있었다. 본 연구에서 개발된 스트립이 자동 분석기기의 재료로 활용될 수 있는지 여부를 알아보기 위하여 분광 비색계로 발색 반응의 최적 파장을 분석하고 최적 파장에서 각 혈당 농도별 반사 밀도를 측정하여 검정선을 얻었다. 이 검정선에 의해 임상 혈청시료의 혈당 농도를 측정한 결과 일본의 Kyoto Daiichi사의 혈액 분석시스템과 유사한 결과를 얻었다. 이로서 본 연구에서 개발한 혈당 스트립을 이용하여 혈액 중 혈당량을 육안으로 측정할 수 있음은 물론 자동 분석기기의 기본 시료로 이용될 수 있음을 확인할 수 있었다.

• 대한수의학회지
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• 제45권2호
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• pp.191-197
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• 2005

An immunochromatographic (IC) strip for the rapid detection of Salmonella spp. in the enriched sample was developed. Affinity purified Salmonella polyclonal antibody was conjugated with 40 nm colloidal gold particles which were prepared by citrate method in our laboratory. The antigen-antibody-gold complex was captured by Salmonella antibody attached to test line of nitrocellulose membrane during the capillary migration of sample. Specificity of the IC strip was calculated to be 100% (12/12) and sensitivity was 97.6% (41/42) in the test with pure cultured bacteria. Salmonella was artificially inoculated into raw pork macerated with enrichment broth. And then it was 10-fold diluted from $5.2{\times}10^{8}CFU/ml$ to 5.2 CFU/ml. The IC strip could detect $5.2{\times}10^{6}CFU/ml$ before enrichment. However, the lowest limit of detection was 5.2 CFU/ml after overnight incubation. The results indicated that the IC assay was a rapid, economical and simple method with high specificity and sensitivity for the detection of Salmonella spp. without using any equipment.

• Parasites, Hosts and Diseases
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• 제49권3호
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• pp.207-212
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• 2011

Rapid serodiagnostic methods for Toxoplasma gondii infection in cats are urgently needed for effective control of transmission routes toward human infections. In this work, 4 recombinant T. gondii antigens (SAG1, SAG2, GRA3, and GRA6) were produced and tested for the development of rapid diagnostic test (RDT). The proteins were expressed in Escherichia coli, affinity-purified, and applied onto the nitrocellulose membrane of the test strip. The recombinant SAG1 (rSAG1) showed the strongest antigenic activity and highest specificity among them. We also performed clinical evaluation of the rSAG1-loaded RDT in 182 cat sera (55 household and 127 stray cats). The kit showed 0.88 of kappa value comparing with a commercialized ELISA kit, which indicated a significant correlation between rSAG1-loaded RDT and the ELISA kit. The overall sensitivity and specificity of the RDT were 100% (23/23) and 99.4% (158/159), respectively. The rSAG1-loaded RDT is rapid, easy to use, and highly accurate. Thus, it would be a suitable diagnostic tool for rapid detection of antibodies in T. gondii-infected cats under field conditions.