• Title/Summary/Keyword: Multiplex

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Development of a Multiplex PCR for Discrimination of the TLC:RS1:CTX array of Vibrio cholerae Wave 3 El Tor Strains

  • Kim, Eun Jin;Yu, Hyun Jin;Nair, G. Balakrish;Kim, Dong Wook
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2199-2205
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    • 2016
  • Vibrio cholerae O1 serogroup Wave 3 El Tor strains are presently prevalent worldwide. The Wave 3 El Tor strains contain a TLC:RS1:CTX array on chromosome 1, and no element is integrated on chromosome 2. A multiplex PCR optimized to identify the TLC:RS1:CTX array of Wave 3 strains has been developed in this study. By using eight primers, the multiplex PCR can identify the characteristic CTX and RS1 array of Wave 3 strains from various arrays of strains belonging to other Waves. The four amplified DNA fragments of Wave 3 strains have been cloned in a vector, which could be used as a positive control for the multiplex PCR. This multiplex PCR and the positive control set could be useful tools for rapid recognition of Wave 3 El Tor strains.

Detection of Brucella spp. and Leptospira interrogans in the Canine Blood by Multiplex Nested PCR (개 혈액에서 Multiplex Nested PCR기법을 이용한 Brucella spp. 및 Leptospira interrogans 검출)

  • Lee, Jung-Youn;Lee, Sang-Eun;Kim, Suk;Kim, Duck-Hwan;Song, Kun-Ho
    • Journal of Veterinary Clinics
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    • v.25 no.4
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    • pp.241-244
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    • 2008
  • This study examined the prevalence of Brucella spp. and Leptospira interrogans in 360 clinically healthy dogs using multiplex nested PCR. Four dogs (1.1%, 2 females and 2 males) tested positive to Brucella spp. by multiplex nested PCR. Fifty nine (16.4%, 31 females and 28 males) of 360 dogs tested positive L. interrogans. In 1 and 2 of the samples that tested positive to Brucella spp. and L. interrogans, the partial sequences of the virB1 and 16S rRNA genes were identified by direct sequence analysis, respectively. In conclusion, prevalence of Brucella spp. and L. interrogans by multiplex nested PCR revealed low and high, respectively. Multiplex nested PCR is can be useful for early detection of Brucella spp. and L. interrogans in the canine blood from asymptomatic dogs.

Multiplex Real-Time PCR for Simultaneous Detection of 6 Periodontopathic Bacteria (Multiplex Real-Time PCR을 이용하여 6종의 주요 잇몸질환 유발 미생물을 동시에 검출하는 기법)

  • Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.292-296
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    • 2013
  • This study utilized an analysis method for detecting six microorganisms, such as Actinobacillus actinomycetemcomitans, Campylobacter rectus, Porphyromonas gingivalis, Tannerella forsythus, Treponema denticola, and Prevotella intermedia, triggering periodontal disease, using multiplex real-time polymerase chain reaction (PCR). The analysis including internal control was made by dividing the six species into two groups using four fluorescence dyes, and it was verified that there was no interference or cross-reaction between the target species and different kinds of oral microbial species. Qualitative and quantitative analyses were conducted on each microorganism in various samples, such as saliva and the plaque, using the multiplex real-time PCR and comparative analysis between periodontitis patients and healthy people, revealing obvious differences between them.

A Comparison between Korean and Chinese Consumers in Service Quality Evaluation: Focused on the Multiplex Cinema

  • LEE, Seung-In;PARK, Yoonseo;JIN, Yanchun;ZHANG, Yan
    • Journal of Distribution Science
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    • v.18 no.8
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    • pp.89-102
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    • 2020
  • Purpose: Our purpose in this study is to compare the SERVQUAL, SERVPERF, and non-difference score measures and to find out which one is better for measuring the service quality of the multiplex cinema service. We also aim to analyze the structural relationships between service quality, customer satisfaction and customer loyalty. Methodology: For the study, we collected data from respondents who have used the multiplex cinema services and conducted an empirical test. SPSS 18.0 was used for descriptive frequency analysis, reliability analysis, and multiple regression analysis, AMOS 18.0 was used for structural equation modeling analysis of causal relationships among variables introduced in research hypotheses. Results: The main results of this study are as follows. First, we found that the non-difference score measure provided a much better model than did other service-measuring models (SERVQUAL, SERVPERF) in Korean and Chinese multiplex cinema. Second, two service-quality factors (Korea-tangibles and assurance vs. China-tangibles and empathy) between the multiplex cinema service quality factors significantly influenced customer satisfaction, which had a significant effect on customer loyalty in Korean and Chinese multiplex cinema. Conclusions: Based on the results, the authors discuss the implications and limitations of this study and future research directions at the end of the paper.

Chemoprevention Effect of Polyozellus multiplex, a Wild and Edible Mushroom (야생식용버섯인 까치버섯(Polyozellus multiplex)의 암예방 효과)

  • Kim, Hyun-Jeong;Han, Jung;Yang, Eui-Joo;Lee, Kap-Rang;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.161-167
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    • 2000
  • Methanol extract and its fractions of Polyozellus multiplex inhibited the growth of several tumor cell lines and its water fraction showed a higher cytotoxicity effect on the human gastric carcinoma cell, SNU668 than on the other cell lines. The glutathione S-transferase (GSH) content was decreased by MNNG treatment but increased by adding Polyozellus multiplex water fractions. Also the activities of GSH and superoxide dismutase were increased by more the treatment of Polyozellus multiplex water fractions than by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) alone. Polyzellus multiplex water fraction cased significant reduction in the proliferating cell unclear antigen (PCNA) labelling index in the glandular stomach epithelium as compared with the value of MNNG alone.

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Multiplex Reverse Transcription-PCR for Simultaneous Detection of Reovirus, Bovine Viral Diarrhea Virus, and Bovine Parainfluenza Virus during the Manufacture of Cell Culture-derived Biopharmaceuticals (세포배양 유래 생물의약품 제조공정에서 Reovirus, Bovine Viral Diarrhea Virus, Bovine Parainfluenza Virus 동시 검출을 위한 Multiplex Reverse Transcription-PCR)

  • Oh, Seon Hwan;Bae, Jung Eun;Kim, In Seop
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.339-347
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    • 2012
  • Viral safety is an important prerequisite for clinical preparations of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacturing process. In particular, Chinese hamster ovary (CHO) cells are highly susceptible to several RNA viruses including reovirus (Reo), bovine viral diarrhea virus (BVDV), and bovine parainfluenza virus (BPIV) and there have been reports of such viral contaminations. Therefore, viral detection during the CHO cell process is necessary to ensure the safety of biopharmaceuticals against viruses. In this study, a multiplex reverse transcription (RT)-PCR assay was developed and subsequently evaluated for its effectiveness as a means to simultaneously detect Reo, BVDV, and BPIV during the manufacture of cell culture-derived biopharmaceuticals. Specific primers for Reo, BVDV, and BPIV were selected, and a multiplex RT-PCR was optimized. The sensitivity of the assay for simultaneous amplification of all viral target RNAs was $7.76{\times}10^2\;TCID_{50}/ml$ for Reo, $7.44{\times}10^1\;TCID_{50}/ml$ for BVDV, and $6.75{\times}10^1\;TCID_{50}/ml$ for BPIV. The multiplex RT-PCR was proven to be very specific to Reo, BVDV, and BPIV and was subsequently applied to the validation of CHO cells artificially infected with each virus. It could detect each viral RNA from CHO cells as well as culture supernatants. Therefore, it was concluded that the multiplex RT-PCR assay can be applied to detection of the adventitious viruses during the manufacture of cell culture-derived biopharmaceuticals.

Comparison of Multiplex Latex Bead Flow Cytometric Analysis and Quellung Analysis in Serotyping Pneumococci (Multiplex Latex Bead 유세포 분석기 기법과 Quellung 반응을 이용한 폐구균 혈청형 분석 비교)

  • Choi, Kyong Min;Yeon, Soo In;Kim, Eun Sook;Shin, Jeon Soo;Yong, Dong Eun;Lee, Kyoung Won;Kim, Dong Soo
    • Pediatric Infection and Vaccine
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    • v.13 no.2
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    • pp.99-105
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    • 2006
  • Purpose : The serotyping results of the Quellung reaction was compared with the newly developed multiplex latex assay and the sensitivity of the Quellung reaction was compared. Methods : We checked the serotypes of 35 samples obtained from patients treated at Yonsei University Medical Center using the multiplex latex bead method and compared the results with the serotypes previously obtained via the Quellung reaction. Results : A decrease in the mean fluorescence was detected in the samples tested with the multiplex assay. Seventeen samples out of the 27 samples agreed to the results of the Quellung assay. We were only able to confirm the concordance of 11 serotypes out of 14 serotypes available. Conclusion : The Quellung reaction is time consuming procedure and prone to errors even with expertise in the procedure, and other alternate methods in serotyping have been investigated to overcome these problems. The newly developed multiplex latex bead assay can test more samples at the same time and has a higher degree of sensitivity. A large scale trial is required to test the sensitivity of the new assay across various serotypes along with efforts to increase the sensitivity of the Quellung assay. The preliminary data suggests that this method may be widely used.

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Development of Raw Material Identification Method of Changnan-jeot and Gaiyang-jeot Using Multiplex PCR and Real-Time PCR (Multiplex PCR과 Real-Time PCR을 이용한 창난젓과 가이양젓 원료 검사법 개발)

  • Choi, Seong Seok;Seo, Yong Bae;Kim, Jong-Oh;Yang, Ji-Young;Shin, Jiyoung;Kim, Gun-Do
    • Journal of Food Hygiene and Safety
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    • v.36 no.4
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    • pp.289-297
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    • 2021
  • In this study, multiplex PCR and real-time PCR were performed on Theragra chalcogramma (walleye pollock), Pangasianodon hypophthalmus (iridescent shark) and their processed foods, such as changnan-jeot and gaiyang-jeot (salted iridescent shark intestine). Species-specific primers for T. chalcogramma and P. hypophthalmus were designed, and genomic DNA was directly extracted from each sample to perform single PCR and multiplex PCR. As a result of PCR, in the case of single PCR, PCR bands of T. chalcogramma (297 bp) and P. hypophthalmus (132 bp) were identified, and in the case of multiplex PCR, it was confirmed that amplification occurred without cross-reaction between T. chalcogramma and P. hypophthalmus. As a result of checking the PCR sensitivity, the concentration of genomic DNA was detected up to 0.1 ng/µL in both single PCR and multiplex PCR. The real-time PCR results showed that the average Ct value of T. chalcogramma was 20.765±0.691, and the average Ct value of P. hypophthalmus sample was 35.719±1.828 in the T. chalcogramma species-specific primers. In the P. hypophthalmus species-specific primers, the average Ct value of the T. chalcogramma sample was 35.996±1.423, and the mean Ct value of the P. hypophthalmus sample was 20.096±0.793. These results demonstrated the significant differences in the efficiency, specificity and cross-reactivity of species-specific primers in real-time PCR. Based on these findings, 7 of changnan-jeot or gaiyang-jeot products were confirmed by multiplex PCR and real-time PCR, and valid results were confirmed in all samples.

Comparison between Bacterial Culture Method and Multiplex PCR for Identification of Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans from the Dental Plaques (치면세균막내의 Fusobacterium nucleatum과 Actinobacillus actinomycetemcomitans의 동정을 위한 세균배양법 및 Multiplex PCR법의 비교)

  • Kim, Hwa-Sook;Lim, Sun-A
    • Journal of dental hygiene science
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    • v.9 no.2
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    • pp.249-255
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    • 2009
  • This study was carried out for the purpose of comparing bacterial culture method, single PCR, and multiplex PCR for identification of F. nucleatum and A. actinomycetemcomitans in subgingival plaque of adult periodontitis. Targeting 20 patients with adult periodontitis, the subgingival plaque was collected in teeth, respectively, for #16, #36, #44. A bacillus was cultivated by painting it over the solid selective media of F. nucleatum and A. actinomycetemcomitans. Bacterial species were detected in 0 tooth with 12 pieces, respectively. Through single PCR and multiplex PCR, the positive reaction was indicated in 43 teeth with 45 pieces, respectively, as for F. nucleatum, and in 1 tooth with 4 pieces, respectively, as for A. actinomycetemcomitans. In the comparative analysis between bacterial identification methods. F. nucleatum showed the more statistically significant difference(p=0.0(0) in comparison between single PCR and multiplex PCR. Even A. actinomycetemcomitans was indicated significantly(p=0.067) in a case that is based on 0.1 in significant level in the comparison between single PCR and multiplex PCR. In conclusion, as a result of comparing the bacterial identification methods, the detection frequency was indicated to be higher in PCR than in bacterial culture method. Single PCR and multiplex PCR showed the mutually similar detection frequency. Accordingly, given thinking of economic efficiency, quickness, and reduction in labor force, it is thought to be more efficient method to use single PCR as the bacterial identification method.

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A Optimization of Fire Safety Design in Multiplex Cinema Theater (국내 복합상영관 시설의 방재대책에 관한 연구)

  • 허준호;윤명오
    • Fire Science and Engineering
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    • v.16 no.2
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    • pp.43-50
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    • 2002
  • 21C, Multiplex Cinema Theater is made from movie industry developed and become a place where we can spend our free time and consuming. But, theater users, who are many and unspecified persons, are short of information on the Multiplex Cinema Theater, so that they may be injured or killed at the occurrence of fire accident. Particularly, Multiplex Cinema Theater is a place that underground space or top floor in highrise building had many people in theaters. However, this problem is difficult to solve that to protect human being and properties at the occurrence of fire accident in the Multiplex Cinema Theater, In order to overcome such problems, Multiplex Cinema Theater need fire protection solutions facilities concerning particulars to pay attention to the safety on a fire and evacuation. This study analysed Multiplex Cinema Theater concept and computer simulation for risk point.