• Journal of Microbiology and Biotechnology
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• 제34권3호
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• pp.681-688
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• 2024

The accurate and rapid detection of methicillin-resistance of Staphylococcus aureus (SA) holds significant clinical importance. However, the methicillin-resistance detection strategies commonly require complicated cell lysis and gene extraction. Herein, we devised a novel colorimetric approach for the sensitive and accurate identification of methicillin-resistance of SA by combining allosteric probe-based target recognition with self-primer elongation-based target recycling. The PBP2a aptamer in the allosteric probe successfully identified the target MRSA, leading to the initiation of self-primer elongation based-cascade signal amplification. The peroxidase-like hemin/G-quadruplex undergo an isothermal autonomous process that effectively catalyzes the oxidation of ABTS2- and produces a distinct blue color, enabling the visual identification of MRSA at low concentrations. The method offers a shorter duration for bacteria cultivation compared to traditional susceptibility testing methods, as well as simplified manual procedures for gene analysis. The overall amplification time for this test is 60 min, and it has a detection limit of 3 CFU/ml. In addition, the approach has exceptional selectivity and reproducibility, demonstrating commendable performance when tested with real samples. Due to its advantages, this colorimetric assay exhibits considerable potential for integration into a sensor kit, thereby offering a viable and convenient alternative for the prompt and on-site detection of MRSA in patients with skin and soft tissue infections.

• 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.

• Molecules and Cells
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• 제45권7호
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• pp.502-511
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• 2022

Bacterial volatile compounds (BVCs) exert beneficial effects on plant protection both directly and indirectly. Although BVCs have been detected in vitro, their detection in situ remains challenging. The purpose of this study was to investigate the possibility of BVCs detection under in situ condition and estimate the potentials of in situ BVC to plants at below detection limit. We developed a method for detecting BVCs released by the soil bacteria Bacillus velezensis strain GB03 and Streptomyces griseus strain S4-7 in situ using solid-phase microextraction coupled with gas chromatography-mass spectrometry (SPME-GC-MS). Additionally, we evaluated the BVC detection limit in the rhizosphere and induction of systemic immune response in tomato plants grown in the greenhouse. Two signature BVCs, 2-nonanone and caryolan-1-ol, of GB03 and S4-7 respectively were successfully detected using the soil-vial system. However, these BVCs could not be detected in the rhizosphere pretreated with strains GB03 and S4-7. The detection limit of 2-nonanone in the tomato rhizosphere was 1 µM. Unexpectedly, drench application of 2-nonanone at 10 nM concentration, which is below its detection limit, protected tomato seedlings against Pseudomonas syringae pv. tomato. Our finding highlights that BVCs, including 2-nonanone, released by a soil bacterium are functional even when present at a concentration below the detection limit of SPME-GC-MS.

• 한국식품과학회지
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• 제36권6호
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• pp.1020-1025
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• 2004

생식제품 및 제조 공정 중 위해 미생물의 제어 및 저감화를 위하여 제조공정을 개선하고 전해수, 오존수 등 살균효과에 대하여 조사하였다. 7가지의 위해미생물 오염도가 높은 생식원료에 대한 전해수의 살균효과가 차아염소산소다수 또는 오존수에 비하여 전체적으로 강한 것으로 나타났으며 오존수의 살균효과는 차아염소산소다수와 비슷하거나 다소 강한 것으로 나타났다. 그러나 전해수나 오존수를 이용하여 생식원료를 세정, 살균하여도 원료의 종류에 따라 살균효과에 차이가 있으며, 비 가열가공이라는 생식의 특성상 원료에서 검출되는 위해미생물은 최종제품까지 생잔할 가능성이 높기 때문에, 각 원료에 따른 살균처리지침을 위한 database 축적이 요구된다. 한편 모델공장을 선택하여 공장위생, 종업원 개인위생 및 제조공정을 개선한 후 공정개선의 미생물학적 효과를 조사한 결과 공정개선 전에는 원료에서 보다 최종제품에서 위해미생물의 수가 증가하는 경향을 나타내었으나, 공정개선 후에는 원료의 균수를 그대로 유지하거나, 감소하였으며 외주 원료의 혼합으로 인하여 위해미생물의 균수가 증가하였다. 따라서 생식의 제조공정에서 위해미생물을 저감화를 위해서는 외주 원료의 철저한 위생관리가 요구되며, 전해수 오존수 등을 이용한 원료의 비가열 세정살균과 지속적이고 위생적인 공정관리가 이루어진다면 위해 미생물의 균수를 저감화할 수 있을 것으로 사료된다.

• 생명과학회지
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• 제10권6호
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• pp.630-639
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• 2000

The gastric pathogen Helicobacter pylori(H. pylori) establishes long-term chronic infection that can lead to atrophic gastritis, intestinal metaplasia, and gastric cancer. H. pylori, which express cytotoxic genes is now recohnized as a cause of peptic ulcer and is also a major risk factor for the development of gastric adenocarcinoma. We performed this study 1) to assess the detection rate of H. pylori according to direct investigation of bacteria of gastric biopsy specimen and two serologic tests of GAP test and Helico blot 2.0 system in the symptomatic and non-symptomatic group 2) to evaluate and compare the efficacy of two serologic tests of GAP test and Helico blot 2.0 system for the diagnosis of H. pylori infection. Forty-nine patients were positive for H pylori infection based on direct investigation of bacteria by histology. The detection rates of H. pylori infection based on direct investigation of bacteria by histology. The detection rates of H. phlori were significantly lower in gastric cancer than in other gastroduodenal disease(p<0.05). The concordance of two serologic tests of GAP test and Helico blot 2.0 system is poor. There was no statistically significant difference between the expression rate of CagA and VacA in the symptomatic and non-symptomatic group. Although Helico blot 2.0 system may not displace GAP test, it was a very sensitive serologic test for the diagnosis of H. pylori infection and it was used to detect IgG antibodies to H. pylori-specific antigens, including CagA, VacA and the various urease subunit. Our data suggest that further investigation is needed to determine whether or not the serologic expression of cytotoxic gene may be clinical usefulness of diagnostic methods in the gastroduodenal disease.

• Journal of Microbiology and Biotechnology
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• 제30권7호
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• pp.955-961
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• 2020

Lactic acid bacteria (LAB) have caused many microbiological incidents in the brewing industry, resulting in severe economic loss. Meanwhile, traditional culturing method for detecting LAB are time-consuming for brewers. The present review introduces LAB as spoilage microbes in daily life, with focus on LAB in the brewing industry, targeting at the spoilage mechanism of LAB in brewing industry including the special metabolisms, the exist of the viable but nonculturable (VBNC) state and the hop resistance. At the same time, this review compares the traditional and novel rapid detection methods for these microorganisms which may provide innovative control and detection strategies for preventing alcoholic beverage spoilage, such as improvement of microbiological quality control using advanced culture media or different isothermal amplification methods.

• 한국축산식품학회지
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• 제32권6호
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• pp.706-712
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• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• Restorative Dentistry and Endodontics
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• 제21권1호
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• pp.1-18
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• 1996

There are many advantages when using IIF and DNA probe methods over anaerobic culture method in that they are time-and effort-saving, more precise and more sensitive. Furthermore, in IIF and DNA probe methods, the detection is possible only with small amount of bacteria, the quantitative analysis is possible, and the cell viability is not necessary. The purpose of this study is to observe the incidence of P.endodontalis by carrying out anaerobic culture, IIF and colony lift using DNA probe method respectively, and to compare these 3 methods in terms of effectiveness and sensitivity in order to identify the most effective detection method. 30 teeth with at least one clinical symptoms, with single canal, and with pulp necrosis were sampled. For sampling bacteria, access cavity was prepared after disinfecting tooth and its surroundings. Then the paper point was inserted up to the periapical area, leave there for a while, and finally it was placed into PRAS Ringer's sol. and PBS sol. In anaerobic culture method, P.endodontalis was identified by biochemical tests after subculturing black and brown colonies which were produced after 7 days of incubation on BAP and Brucella BAP in anaerobic chamber. To identify P.endodontalis in IIF method, species-specific polyclonal rabbit-antisera of P.endodontalis(ATCC 35406) was reacted with sampled PBS sol. dispensed onto glass slide, and then P.endodontalis was examined by phase contrast microscopy after incubating with Goat anti-rabbit lgG conjugated to Fluorescein isothiocyanate. For colony lift using DNA probe method, membranes were laid over colonies on the surface of BAP and were hybridized with cloned DNA probe of P.endodontalis. The existence of P.endodontalis was then identified by the methods of chemiluminescent detection and color metric detection. Black colony was found in 11 teeth out of 30 teeth and P.endodontalis was detected in 6 teeth (20 %) by anaerobic culture method, 16 teeth (53 %) by IIF method, and 7 teeth (23 %) by DNA probe method. IIF method is significantly better in detecting P.endodontalis than DNA probe method and anaerobic culture method. There was no significant differences between DNA probe method and anaerobic culture method. There was significant correlation between the formation of black colony and the existence of P.endodontalis. The probability of detecting P.endodontalis when black colony being present is 2.89 times higher than when not being present. There was significant relationship between the foul odor of clinical symptoms and P.endodontalis. The sensitivity of existing P.endodontalis when foul odor being present was 93.75 %, while the specificity of not existing P.endodontalis when foul odor not being present was 28.57 %. These results suggested that the probes of P.endodontalis will be used to decide the method and prognosis in endodontic treatments.

• 대한치과보철학회지
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• 제46권2호
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• pp.116-124
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• 2008

문제 진술: 임플랜트 시스템은 임플랜트와 지대주 사이에서 빈 공간이 발생하여 세균의 저장소로서 작용하게 되어 임플랜트 주위 연조직에 염증반응을 야기할 수 있다. 목 적: 이 연구의 목적은 임플랜트-지대주 간극의 미세 누출에 관여하는 치주질환원인균으로 추정되는 Porpyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Treponema denticola, Aggregatibacter actinomycetemcomitans의 검출률을 조사하는 것이다. 연구재료 및 방법: 시료채취는 27명의 환자들에서 행하였고 소독된 페이퍼포인트를 이용하여 $1{\times}PBS$로 옮겼다. 치주질환원인균의 검출은 16 rDNA에 근거한 종특이적인 프라이머를 지닌 중합효소체인반응을 이용하였다. 결과: 임플랜트 고정체내에서의 Porphyromonas gingivalis와 Prevotella intermedia의 검출률은 59%와 82%였다. 환자의 임플랜트 연구에서는 Porphyromonas gingivalis와 Prevotella intermedia의 검출률은 44%와 82%였다. 환자혀에서의 Porphyromonas gingivalis와 Prevotella intermedia의 검출률은 82%와 82%였다. 결 론: 현재의 임플랜트 시스템은 임플랜트 내부의 세균 군집화와 미세누출을 안전하게 예방할 수 없다.

• 한국식품영양학회지
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• 제17권3호
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• pp.254-259
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• 2004

Primer 20가지로 Listeria spp.에 대해 screening을 하여 병원균인 L. monocytogenes를 구별하게 하는 RAPD-PCR의 band pattern을 나타내는 10-mer random primer가 OPG-13이 라는 것을 알았다. OPG-13은 GC%가 70%이므로 계산상으로는 annealing 온도가 34$^{\circ}C$이다. 32～36$^{\circ}C$까지 다섯 가지의 온도로 annealing한 결과 L. monocytogenes만이 갖는 특정한 크기의 band가 역시 34$^{\circ}C$에서 형성됨을 알아냈고, 34$^{\circ}C$를 annealing 온도로 정하였다. Line 1부터 4까지 1. monocytogenes ATCC15313, 19111, 19112, 19113은 2개의 700 bp와 1500 bp band를 형성하였고 그 밖의 Listeria spp.들 Line 6부터 11까지 L. ivanovii ATCC 19119, L. grayi ATCC19120, L. murrayi ATCC25401, L. innocua ATCC33090, L. welshimeri ATCC35897, L. seeligeri ATCC35967은 약 2,000 ～ 2,300 bp크기 한 개의 band를 보여 병원성 균과 비병원성 균이 매우 확실하게 구분되는 band pattern이 나타나는 이러한 결과로 10-mer random primer인 OPG-13을 이용한 RAPD-PCR 방법이 병윈균인 L. monocytogenes를 검색하는데 이용될 수 있음을 확인할 수 있었다.