• Korean Journal of Food Science and Technology
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• v.45 no.1
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• pp.133-136
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• 2013

The purpose of this study was to compare a conventional culture method and real-time PCR for the detection of Yersinia enterocolitica (Y. enterocolitica) in sausage and in vegetable salad. Food samples inoculated with Y. enterocolitica were enriched in peptone-sorbitol bile-broth, and swabs were then streaked onto cefsulodin-irgasan-novobiocin agar. Biochemical tests for suspected colonies were performed with an API 20E strip. In parallel, real-time PCR was performed, targeting the 16S rRNA gene using 1 mL of enrichment broth. In sausage, the number of positive samples detected by culture method (49 out of 60) was similar (p>0.05) with that of real-time PCR (50 out of 60). However, the number of positive samples of real-time PCR (26 out of 60) was significantly higher (p<0.05) than that of the conventional culture method (6 out of 60) in vegetable salad. Real-time PCR could be an effective screening tool for detecting Y. enterocolitica, particularly in food samples with high levels of background flora, such as a vegetable salad.

• Tuberculosis and Respiratory Diseases
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• v.85 no.3
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• pp.264-272
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• 2022

Background: The current conventional drug susceptibility test (DST) for Mycobacterium tuberculosis (Mtb) takes several weeks of incubation to obtain results. As a rapid method, molecular DST requires only a few days to get the results but does not fully cover the phenotypic resistance. A new rapid method based on the ability of viable Mtb bacilli to hydrolyze fluorescein diacetate to free fluorescein with detection of fluorescent mycobacteria by flow cytometric analysis, was recently developed. Methods: To evaluate this cytometric method, we tested 39 clinical isolates which were susceptible or resistant to isoniazid (INH) or rifampin (RIF), or ethambutol (EMB) by phenotypic or molecular DST methods and compared the results. Results: The susceptibility was determined by measuring the viability rate of Mtb and all the isolates which were tested with INH, RIF, and EMB showed susceptibility results concordant with those by the phenotypic solid and liquid media methods. The isolates having no mutations in the molecular DST but resistance in the conventional phenotypic DST were also resistant in this cytometric method. These results suggest that the flow cytometric DST method is faster than conventional agar phenotypic DST and may complement the results of molecular DST. Conclusion: In conclusion, the cytometric method could provide quick and more accurate information that would help clinicians to choose more effective drugs.

• The Journal of the Korean Society for Microbiology
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• v.21 no.3
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• pp.381-385
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• 1986

Twenty strains of penicillin(PC)-resistant Staphylococcus aureus ($MIC{\geqq}32U/ml$) were chosen randomly from recent isolates and submitted to the present experiment to see what effect the combined antibiotic action of moxalactam(MX) and fosfomycin(FM) would bring about on the cells, using MS-2 system. 1. The conventional agar dilution method and the rapid automatic MS-2 system were used in measuring the MICs of MX and FM against each strain and the comparison of the data obtained revealed no significant difference between the two methods in the titer and distribution of the MICs. 2. The automatic MS-2 system was, therefore, used alone in determining the combined growth inhibitory effect of MX and FM because of its more rapidness, and the obtained results were that most of the PC-resistant strains(16 out of 20, 80%) were synergistically inhibited by the two antibiotic combination while additive effect was observed in the remaining 4 strains(20%). 3. Thus, it is suggested that the growth of PC-resistant staphylococcal cells may be synergistically inhibited by MX and FM combination and the efficiency of two antibiotic action as well as MIC of single antibiotic may be more rapidly determined by the MS-2 system than by the conventional method.

• Journal of Veterinary Clinics
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• v.27 no.4
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• pp.402-406
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• 2010

Bovine leukemia virus (BLV) is a delta-retrovirus which causes chronic lymphocytosis in cattle. BLV infections have been divided into two groups such as enzootic bovine leukosis (EBL) and sporadic bovine leukosis (SBL) according to the clinical symptoms in infected cattle. The conventional detection method of BLV was hematological procedure which is determining lymphocytosis in the suspected animals. Recently several sensitive methods were developed to detect antibody to BLV and nucleic acid of the BLV from infected cattle. In this study we have compared the difference of positive rates between agar gel immunodiffusion (AGID) and enzyme linked immunosorbent assay (ELISA) which are using for BLV antibody detection methods. The positive detection rate of ELISA test was 7.4% greater than the positive rate of AGID. The discrepancy of the positive rate between ELISA and AGID were showed in the group of age over one year old to under three year old group. The result from each test agreed very well in the group of over 5 year old cattles. The serological test is very useful method to select the infected cattle for the eradication or control of the disease in the infected herd. But it has a limit by interference of the maternal antibody from the cow of under 6 month old. This study shows that 16.2% of these ages group showed BLV gene positive by polymerase chain reaction (PCR) method. The result suggests that ELISA test need to be used with PCR to clarify misinterpretation of positive animals by antibody response due to the natural infection from maternally derived antibody in calves of under 6 months old.

• Journal of Microbiology
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• v.45 no.6
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• pp.515-520
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• 2007

Vaginal lactic acid-producing bacteria of 80 pre-menopausal women were studied by isolation on Blood and DeMan-Rogosa-Sharpe agar, PCR with group-specific primers for Lactobacillus-denaturing gradient gel electrophoresis (DGGE), and PCR with specific primers for V3 region in 16S rRNA-temporal temperature gel electrophoresis (TTGE). Conventional isolation method on media detected only one lactobacillus (Lactobacillus brevis) while TTGE detected only Lactobacillus sp. DGGE detected seven Lactobacillus species; L. coleohominis, L. crispatus, L. iners, L. reuteri, L. rhamnosus, L. vaginalis, and Leuconostoc lactis. L. acidophilus and L. gasseri, which are prevalent in Western women, were not detected in Korean women. Furthermore, L. rhamnosus, Leuc. lactis, L. coleohominis, and Weissella cibaria, which were not previously reported in the vaginal microbiota of Korean women, were detected. The five most prevalent LABs in vaginal microbiota in Korean women were L. iners, Enterococcus faecalis, L. crispatus, Leuc. lactis, and W. cibaria.

• Korean Journal of Microbiology
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• v.18 no.3
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• pp.148-152
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• 1980

The microwave of 2450 MHz, generated by a household cooking oven, was evaluated for its applicability to melt various rehydrated media and to remove dissolved oxygen from tubed media for anaerobic culture. The effect on the sterilization of E. coli in selective media was also evaluated. The following results were obtained. 10 The microwave oven was useful in saving time for melting media and in eliminating heat and combustion gas from the laboratory, which were inevitable by-products in the conventional flame method. 2) Dissolved oxygen could be removed without boiling over by exposing the tubes of anaerobic culture medium after putting them in a wire basket in a beaker with water. 30 The count of E. coli during the melting of MacConkey and EMB agar were similar to those treated with open flame. The microwave treatment was not considered a possible mean to replace autoclaving even in these selective media.

• Journal of Food Hygiene and Safety
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• v.34 no.3
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• pp.309-315
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• 2019

To enumerate Staphylococcus aureus in food, Baird-Parker Agar (BPA) is usually used in the conventional method, However it requires time and space for the preparation and plating, and incubation. Thus, use of the $3M^{TM}$ $Petrifilm^{TM}$ Staph Express Count Plate (STX Petrifilm) might be appropriate to solve these challenging problems. The purpose of this study was to compare the efficiency of STX Petrifilm with BPA for enumeration of S. aureus in various foods. A mixture of S. aureus strains ATCC29213, ATCC25923, and ATCC13565 was inoculated on marinated pork chop, beef (chuck tender), dried filefish, semi-dried squid, rice cake, and Japchae (stir-fried glass noodles) at 2, 3, 5, and 7 Log CFU/g. S. aureus cell counts were enumerated by spread-plating on STX Petrifilm and BPA after 0 and 24 hours at $4^{\circ}C$ (marinated pork chop, beef, semi-dried squid, and stir-fried glass noodles) and $25^{\circ}C$ (dried filefish and rice cake). Recovery of STX Petrifilm for S. aureus from various food samples was compared with BPA, and the results showed that there were no significant differences between two selective media in all cases. The results indicated that STX Petrifilm had enough efficiency to recover S. aureus from various foods as well as saving time and space.

• Korean Journal of Microbiology
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• v.47 no.1
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• pp.87-91
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• 2011

The aim of this study was to compare the performance of conventional culture and real-time PCR for detection of Cronobacter spp. in powdered foods. Infant formula, baby food and Misugaru inoculated with Cronobacter were enriched in distilled water as first enrichment step, followed by incubating in Enterobacteriaceae enrichment (EE) broth as second enrichment step. A loopful of enriched sample was streaked onto Druggan-Forsythe-Iversen agar, followed by incubating at $37^{\circ}C$ for 24 h. One milliliter of the enriched distilled water and EE broth were used in real-time PCR assay. No statistical differences were observed in the number of positive samples between culture method and real-time PCR (p>0.05) in all types of food samples. The number of positives of real-time PCR was higher in the first enrichment media (distilled water) than the second enrichment media (EE broth), though there was no significant difference (p>0.05). It appears that some components of the second enrichment broth, EE broth, inhibit the reaction of real-time PCR. These results show that real-time PCR using a single enrichment with distilled water could be useful as an effective screening method for detection of Cronobacter while saving much time and labor compared to conventional culture method.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.2
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• pp.177-184
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• 2019

Mycobacteria grow slowly. Therefore, a solid medium should be used for eight weeks and a liquid medium for six weeks. The purpose of this study was to find the growth factors that can grow Mycobacterium rapidly and to help develop a solid medium for rapid identification. Three types of Mycobacterium growth factors were evaluated with 10 Mycobacteria by adding activated charcoal, defibrinated sheep blood, and L-ascorbic acid to $Difco^{TM}$ Mycobacteria 7H11 agar (Becton, Dickinson and Company, Sparks, MD, USA). The time to detection and the distinguishability of a colony were compared with that of the current method. In the rapidly growing Mycobacterium, the difference in detection time between the new media and conventional media confirmed that the new media was faster. M. kansasii and M. intracelluare grew faster in 7H11 C than in 7H11 medium. MTB grew faster than the other media in 7H11 C. This study confirmed that the two growth factors affect fast-growing Mycobacteria and slow-growing Mycobacteria. 7H11 C showed better distinguishability than the conventional media in all 10 Mycobacterium due to the color contrast. In particular, when the MTB was grown, the size of the colonies was larger than with other media, so visualization was easy.

• Journal of Microbiology and Biotechnology
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• v.5 no.2
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• pp.80-86
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• 1995

The bacterial strain WC-17 able to produce chitinase was isolated from soil using an enrichment technique. The isolated strain was identified as Acinetobacter sp. judging by their morphological and physiological characterisitics. The optimal culture conditions for the production of chitinase of Acinetobacter sp. WC -17 are 1.5% colloidal chitin and 1 % tryptone at $30^{\circ}C$ with pH 6.5. Since the enzyme was rapidly produced in a culture supplied with chitin, glucose, or N-acetylglucosamine but not with other polymers and monosaccharide, the enzyme was considered to be an inducible enzyme. Notably N- acetylglucosamine and glucose were found to be effective inducers at low concentrations but repressors at excessive concentrations. The cultural supernatant of Acinetobacter sp. WC-17 inhibited the growth of phytopathogenic fungi such as P.oryzae, R.solani, and F.solani. Among the phytopathogenic fungi tested, P.oryzae was the most sensitive. The conventional agar plate (PDA containing 1 % colloidal chitin) method also produced the same result.