• Journal of Microbiology and Biotechnology
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• 제19권1호
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• pp.83-92
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• 2009

Individual immunochromatographic assays (ICG) for ochratoxin A (OTA) and zearalenone (ZEA) were optimized and used in the development of a one-step simultaneous immunochromatographic assay (OS-ICG) for the rapid multianalysis of two mycotoxins in corn samples. The nitrocellulose membrane of the OS-ICG was treated with OTA-bovine serum albumin (BSA), ZEA-ovalbumin (OVA), and anti-mouse IgG in the OTA test, ZEA test, and control zones, respectively. Monoclonal antibody-gold conjugates (OTA3 MAb-gold and ZEA2C5 MAb-gold) were sprayed onto the conjugate pad. The visual detection limits were 2.5 and 5 ng/ml for OTA and ZEA, respectively, and the results were obtained within 15 min after starting the analysis. An efficient, simple, and rapid extraction method using 30% MeOH/PBS was established and validated by analyzing the corn samples spiked with OTA/ZEA mixtures (0/0, 5/10, 10/20, and $20/30\;{\mu}g/kg$). The cut-off values of the OS-ICG for the spiked corn were 5 and $10\;{\mu}g/kg$ for OTA and ZEA, respectively. Natural corn samples were analyzed by OS-ICG, direct competitive enzyme-linked immunosorbent assay (DC-ELISA), and HPLC. Results of the OS-ICG were in good agreement with those obtained by DC-ELISA and HPLC. The developed OS-ICG offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site simultaneous determination of OTA and ZEA in cereals, food, and agricultural products in one analytical cycle.

• Food Science and Biotechnology
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• 제18권3호
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• pp.641-648
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• 2009

A one-step simultaneous immunchromatographic (OS-ICG) assay using colloidal gold-monoclonal antibody (gold-MAb) conjugates was developed for the rapid multianalysis of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in feed samples. Visual detection limits for AFB1 and OTA were 0.5 and 2.5 ng/mL, respectively, and the results were obtained within 15 min. Matrix interference from the feed extracts was efficiently reduced by appropriate dilution with buffer. Cut-off values of the OS-ICG assay for the feed spiked with AFB1/OTA mixtures (5/5, 10/10, 25/25, 50/55, 100/100 ${\mu}g/kg$) were 10 and 50 ${\mu}g/kg$ for AFB1 and OTA. The comparative analyses of 65 feed samples by OS-ICG, enzyme-linked immunosorbent assay (ELISA), and high performance liquid chromatography (HPLC) showed good agreement. In this study, we confirmed that simultaneous analysis based on immunoassay is possible and it can be used as an on-site multianalysis of AFB1 and OTA in feed, food, and agricultural products.

• 대한수의학회지
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• 제45권2호
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• pp.169-177
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• 2005

We developed the one-step strip based on an immunochromatographic (IC) assay for the rapid detection of Listeria spp. Genus-specific monoclonal antibody to flagella of L. monocytogenes was conjugated with 40 nm colloidal gold particles which were prepared in our laboratory. The specificity of the IC strip was tested with pure cultured bacteria. All strains of the genus of Listeria spp. yielded positive reactions and 12 strains of non-Listeria were negative, resulting in a specificity of 100%. L. monocytogenes was artificially inoculated in raw pork macerated with listeria enrichment broth. And then it was 10-fold diluted from $8.7{\times}10^6$ to 8.7 CFU/ml. L. monocytogenes could be detected at a minimum of $8.7{\times}10^5CFU/ml$ before enrichment, $8.7{\times}10^2CFU/ml$ after primary enrichment and 8.7 CFU/ml after secondary enrichment, respectively. These results indicated that the IC strip exhibited high specificity and sensitivity in the detection of Listeria spp.

• 한국임상수의학회지
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• 제31권1호
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• pp.1-5
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• 2014

고양이 백혈병 바이러스(feline leukemia virus, FeLV)는 전 세계적으로 고양이의 가장 일반적이고 중요한 전염성 질환 중 하나이며, 다른 어떤 전염병 보다 고양이에 폐사를 일으키는 중요한 전염병이다. 방글라데시에 있어서 이 질병의 이환율은 집에서 기르는 고양이나 집을 나간 야생 고양이에 대한 예방, 관리, 치료 방침을 정하는데 중요한 자료를 제시할 것이다. 이 연구는 신속 면역크로마토그라피 분석법을 이용한 RapiGEN$^{(R)}$ FeLV 항원검사 키트(RapiGEN$^{(R)}$ Inc., 대한민국)를 사용하여 방글라데시 Mymensingh 지역의 FeLV의 감염률을 조사하기 위해 수행되었다. 총 130마리 고양이(23마리 집에서 기르는 고양이와 107마리의 집을 나간 야생 고양이)의 혈액 샘플은 제조 업체의 사용법에 따라 수집 및 검사를 수행하였다. FeLV 감염의 전체 유병률은 1.54%(2마리/130마리)이었으며, 감염률은 1세 미만의 고양이에서는 1.79%(2마리/112마리)로 나타났으며, 1세 이상의 고양이에서는 모두 음성이었다. 수컷과 암컷의 감염률은 각각 1.72%(1마리/58마리), 1.39%(1마리/72마리)로 나타났다. 집에서 기르는 고양이에 있어서는 모두 음성이었으나 집을 나간 야생 고양이에 있어서 감염률은 1.87%로 나타났다. FeLV 양성은 모두 임상증상이 나타나는 고양이에서 발견되었다. 이 연구결과에 있어서는 성별, 소유 및 건강 상태에 따라 유의한 상관관계가 있는 것은 아니었다. 이 연구결과는 방글라데시에서 FeLV 감염률에 대한 처음 보고이며, 이 검사 방법에 사용한 신속 면역크로마토그라피 분석법을 활용한 항원검사 키트는 저렴하고, 빠른 검사로 결과를 얻을 수 있는 장점이 있어서 임상에 적용하기에 효과적인 것으로 판단된다.