• Title/Summary/Keyword: PCR (Polymerase Chain Reaction)

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Epidemics of Ascetic Meningitis in Kyoungsangnamdo from May to August, 1996 (96년도 상반기에 경상남도 중부지방에서 유행한 무균성 뇌막염에 대한 고찰)

  • Kwon, Oh Su;Lee, Kyoung Lim;Kim, Won Youb;Jung, Won Jo;Ma, Sang Hyouk;Lee, Kyu Man
    • Pediatric Infection and Vaccine
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    • v.4 no.1
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    • pp.97-105
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    • 1997
  • Purpose : Aseptic meningitis mainly caused by enterovirus is common in pediatric population especially during summer & fall. Most of pediatric patients restore their health without any complications with proper management. Between May to August of 1996, Masan and surrounding areas of the Kyoungsangnamdo were epidemic areas for the aseptic meningitis. The purpose of this study was to determine causative virus and describe correlation between disease and clinical symptoms in aseptic meningitis patients and those with fever and characteristic rashes without apparent meningitis symptoms. Methods : Between May to August, 1996, 57 patients with high fever and characteristic feature of rashes were reviewed. From 22 cerebrospinal fluid & 57 stool obtained specimens, viral culture and detection of enterovirus RNA were conducted. Collected specimens were kept in $-30^{\circ}C$ environment until sending of specimens to labortory. The virus identified through indirect immunofluorescence. RT-PCR method was used to identify enterovirus RNA in cerebralspinal fluid. Results : 1) One hundred fifty five pediatric patients with viral infection required hospitalization. Disease occurred higher rate in male than female with ratio of 1.94:1. Examined patients' age ranged from 15days old to 15years old. But most of patients(74.8%) were under age of 5years old. The time of occurrence was between May to August of 1996. 2) All patients had high fever and physical symptoms in those patients include headache, vomiting, abdominal pain, diarrhea, and rashes. The rashes observed mainly in patients under age of 4 years and were predominantly commom patients under age of 18 months olds)<0.001). 3) Between sampled patients and non-sampled patients, clinical course was similar. Echovirus type 9 was cultivated in 41 out of 57 cases of collected stool specimens. RT-PCR that used on CSF showed positive results in 10 out of 22 cases. Three cases of positive cultivated of positive results in RT-PCR were echovirus type 9. Conclusions : Echovirus type 9 was thought to be the causative agent of aseptic meningitis that was prevalent throughout mid areas of Kyoungsangnamdo from May to August, 1996. Additionally causative agent that responsible for high fever with rashes without meningitis symptoms also thought to be the same echovirus type 9.

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Identification of Faulty Red Pepper Powder Containing Seasoned Red-pepper Sauce (고추다대기 혼입 불량고춧가루 판별법 개발)

  • Park, Yong-Chjun;Lim, Ji-Young;Kim, Mi-Ra;Park, Young-Eun;Lim, Jan-Di;Hwang, Cho-Rong;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.27 no.2
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    • pp.182-187
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    • 2012
  • In this study, the experimental method has been investigated using molecular biological way to identify raw materials from seasoned red-pepper sauce which is one of the most popular spices in Korea. 6 kinds of seasoned red-pepper sauces were chosen as a sample containing chilli pepper, garlic, onion as a major ingredient and species specific primers were used for the identification of the raw material of processed food. Selected samples were pre-treated to remove salt (samples were washed with distilled water 3~4 times for desalting), after that, to amplify the extracted genes, whole genome amplification (WGA) kit was performed. Afterwards, PCR products were confirmed through the electrophoresis. As a result, 102, 180, 280 bp of specific PCR products were confirmed for each major ingredients such as chilli pepper, garlic, onion. From this study, the gene extraction method was validated for the identification of ingredients from the spices and it would be applied to distinction of low quality chilli pepper powder including seasoned red-pepper sauce illegally.

Exploration of Virulence Markers and Genes of Listeria monocytogenes Isolated from Animal Products (축산물유래 Listeria monocytogenes의 virulence marker 및 gene 조사)

  • Yi, Chul-Hyeon;Song, Hyeon-Ho;Kim, Mi-Ryung;Kang, Ho-Jo;Son, Won-Geun
    • Journal of Food Hygiene and Safety
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    • v.23 no.3
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    • pp.248-256
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    • 2008
  • To investigate the epidemiological characteristics of 68 Listeria monocytogenes isolates, including 11 reference strains and 57 isolates from imported US beef, domestic meats(beef, pork, chicken meat), raw milk, and milk plants. L. monocytogenes was to evaluate the production of virulence proteins, such as hemolysin(LLO) and lecithinase(LCP), the adsorption of Congo red(CRA), and to detect virulence genes using the polymerase chain reaction(PCR). In the study of virulence protein production, 68(100%), 62(91.2%), and 54(79.4%) of the 68 L. monocytogenes strains were positive for LLO production, the LCP test, and the CRA test, respectively, while strains of other species, such as L. innocua, L. gray, L. murrayi, and L. welshimeri, were not. There were no significant differences between L. monocytogenes serotypes and the ability to produce LLO or LCP. L. monocytogenesstrains had very high hemolytic titers(2 to 16 fold), while the other Listeria species, other than L. ivanovii and L. seeligeri, did not. The hemolysin activities of L. monocytogenes, L. ivanovii, and L. seeligeri usually exceeded 1.0 HU/mg, while those of other Listeria spp. were less than 0.04 HU/mg. In the PCR assay, all of the L. monocytogenes strains contained the hlyA, plcA, plcB, inlA, and inlB virulence genes and produced a product of the expected size. In the PCR of the actA gene, the expected 385-bp product was seen in 39(57.4%) L. monocytogenesstrains, while an unexpected 268-bp product was seen in 29(42.6%) strains. Most L. monocytogenes strains isolated from Hanwoo beef produced the 385-bp actA gene product, while strains of imported US beef usually produced the 268-bp actA gene product. By contrast, no virulence gene products were amplified in the other Listeria spp.

The Clinical Features of Pertussis in Infancy (영아 백일해의 임상 양상)

  • Kim, Soon Ju;Kim, Sang-Eun;Kim, Jong-Hyun;Lee, Jung-Hyun;Oh, Jin Hee;Koh, Dae Kyun;Sung, Hwa Young;Yu, Jae-Yon
    • Pediatric Infection and Vaccine
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    • v.16 no.2
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    • pp.167-174
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    • 2009
  • Purpose : Pertussis was very common in the past, but reported cases have dramatically decreased. The improvement of vaccination programs and unreadiness of laboratory confirmation seems to have developed this situation. This study investigated the frequency of pertussis among infants with a paroxysmal cough and compared the clinical characteristics between infants with and without pertussis. Methods : Between June and November 2006, 27 infants admitted to the hospital that were 15-90 days old with a history of a cough for more than seven days were enrolled. The cough was described as: paroxysmal, whooping, and post-tussive vomiting. PCR and cultures for Bordetella pertussis with nasopharyngeal aspirates were obtained. The patients were divided into two groups: (1) the pertussis group that had positive results by PCR or culture; (2) the control group that had negative results by PCR and culture. Clinical and laboratory characteristics were compared between the two groups. Results : Among the 27 cases, five (18.5%) were finally diagnosed with pertussis. Only one out of the five pertussis cases was initially diagnosed with a pertussis-like syndrome on admission. Compared to the group without pertussis, the pertussis group had a significantly higher frequency of: no fever (P =0.043), a paroxysmal cough (P =0.040), cyanosis (P =0.001), non-immunized status for DTaP (P =0.047), normal auscultation (P =0.028), normal chest X-ray findings (P =0.027), high absolute lymphocyte count (P =0.039), and low CRP (P =0.046). The patients with the diagnosis of pertussis had a significantly longer duration of coughing (27.2${\pm}$10.6 vs. 12.6${\pm}$5.6 days, P =0.039). Conclusion : Pertussis should be suspected in any infant with typical symptoms of pertussis in addition to: a persistent cough without fever, accompanied by paroxysms or cyanosis prior to the age of DTaP immunization. Active laboratory confirmation should be carried out to confirm more cases with pertussis.

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분자유전학적인 기술을 이용한 육 감별법

  • Kim, Tae-Heon
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2000.11a
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    • pp.59-75
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    • 2000
  • This study was carried out to develop a DNA marker for identifying between Korean cattle (Hanwoo) and other breeds. First experiment was performed to isolate Hanwoo specific DNA marker at sequence characterized amplified regions (SCARs). Five breeds of cattle including Hanwoo, Holstein, Hereford, Angus and Charolais were represented with the from 8 to 20 individuals. Fourteen primers of 300 arbitrary primers of 10 nucleotides showed reproducible polymorphism across the breeds. An amplified band of 0.9 kb in the primer MG-3 showed the specificity to Holstein breed. And MG-6 and MG-12 detected the Hereford and Hanwoo specific markers at the size of 2.0 kb and 1.0 kb, respectively. A 1.0 kb band of MG-12 was cloned and sequenced. A SCAR primer was designed based on the obtained sequences. It was possible to identify the Hanwoo from Holstein breed. Second experiment was carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no.Y19103). A size of 350 bp was amplified by polymerase chain reaction(PCR), digested with two different restriction enzyme, BsrFI and MspA II, and electrophoresed in 2.5% Metaphore agarose gel for determination of genotypes. Genotype frequencies of Hanwoo were 0.10 in E+e and 0.90 in ee. Allele ED was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. We suggested that SCAR marker and the bovine MC1R gene could be used as a DNA marker for distinguishing beef between Hanwoo and Holstein.

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Expression of Yippee-Like 5 (YPEL5) Gene During Activation of Human Peripheral T Lymphocytes by Immobilized Anti-CD3 (인체 말초혈액의 활성화 과정 중 yippee-like 5 (YPEL5) 유전자의 발현 양상)

  • Jun, Do-Youn;Park, Hye-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1641-1648
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    • 2007
  • Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.

CT Examinations for COVID-19: A Systematic Review of Protocols, Radiation Dose, and Numbers Needed to Diagnose and Predict (COVID-19 진단을 위한 CT 검사: 프로토콜, 방사선량에 대한 체계적 문헌고찰 및 진단을 위한 CT 검사량)

  • Jong Hyuk Lee;Hyunsook Hong;Hyungjin Kim;Chang Hyun Lee;Jin Mo Goo;Soon Ho Yoon
    • Journal of the Korean Society of Radiology
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    • v.82 no.6
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    • pp.1505-1523
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    • 2021
  • Purpose Although chest CT has been discussed as a first-line test for coronavirus disease 2019 (COVID-19), little research has explored the implications of CT exposure in the population. To review chest CT protocols and radiation doses in COVID-19 publications and explore the number needed to diagnose (NND) and the number needed to predict (NNP) if CT is used as a first-line test. Materials and Methods We searched nine highly cited radiology journals to identify studies discussing the CT-based diagnosis of COVID-19 pneumonia. Study-level information on the CT protocol and radiation dose was collected, and the doses were compared with each national diagnostic reference level (DRL). The NND and NNP, which depends on the test positive rate (TPR), were calculated, given a CT sensitivity of 94% (95% confidence interval [CI]: 91%-96%) and specificity of 37% (95% CI: 26%-50%), and applied to the early outbreak in Wuhan, New York, and Italy. Results From 86 studies, the CT protocol and radiation dose were reported in 81 (94.2%) and 17 studies (19.8%), respectively. Low-dose chest CT was used more than twice as often as standard-dose chest CT (39.5% vs.18.6%), while the remaining studies (44.2%) did not provide relevant information. The radiation doses were lower than the national DRLs in 15 of the 17 studies (88.2%) that reported doses. The NND was 3.2 scans (95% CI: 2.2-6.0). The NNPs at TPRs of 50%, 25%, 10%, and 5% were 2.2, 3.6, 8.0, 15.5 scans, respectively. In Wuhan, 35418 (TPR, 58%; 95% CI: 27710-56755) to 44840 (TPR, 38%; 95% CI: 35161-68164) individuals were estimated to have undergone CT examinations to diagnose 17365 patients. During the early surge in New York and Italy, daily NNDs changed up to 5.4 and 10.9 times, respectively, within 10 weeks. Conclusion Low-dose CT protocols were described in less than half of COVID-19 publications, and radiation doses were frequently lacking. The number of populations involved in a first-line diagnostic CT test could vary dynamically according to daily TPR; therefore, caution is required in future planning.

Development of Detection Method for Oilfish (Ruvettus pretiosus and Lepidocybirium flavobrunneum) as a Food Materials not Usable in Foods (식품원료로 사용금지 대상인 기름치 (기름갈치꼬치 및 흑갈치꼬치) 판별법 개발)

  • Park, Yong-Chjun;Kim, Mi-Ra;Jung, Yong-Hyun;Shin, Joon-Ho;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.28 no.1
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    • pp.50-55
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    • 2013
  • Since 1 June 2012, it is prohibited to sell oilfish as a food material but there are still many illegal cases of selling oilfish as if it is tuna or grilled Patagonian toothfish. So it is absolutely crucial to construct the system to distinguish the real food material from oilfish. There are two sorts of oil fish called Ruvettus pretiosus and Lepidocybirium flavobrunneum involved in Percifomes order and Gempylidae class. 16S DNA gene region in mitochondria was selected to design the specific primers. For design species-specific primer, the theoretical experiment were performed for the sequences of R. pretiosus, L. flavobrunneum, Thunnus thynnus, Thunnus albacores, Makaira mitsukurii and Xiphias gladius, registered at the Gene bank from the National Centre for Biotechnology Information, using BioEdit 7.0.9.0. program. Through the analysis of the result from experiments, it was possible to design the 4 kinds of primers to distinguish R. pretiosus and L. flavobrunneum. As a comparison group, 3 kinds of tuna and 4 kinds of billfishes were selected and experimental verification was performed. As a result, for R. pretiosus and L. flavobrunneum, R.P-16S-006-F/R.P-16S-008-R and L.F-16S-004-F/L.F-16S-006-R primers were selected eventually and PCR condition was established. In addition, 178bp and 238bp of PCR products were confirmed from the established condition and non-specific band was not amplified among similar species. Therefore, the species-specific primers developed in this study would be very useful and used in various ways such as internet shopping mall and illegal distributions with fast and scientific results.

Clinical significance of codetection of the causative agents for acute respiratory tract infection in hospitalized children (급성 호흡기 감염으로 입원한 소아에서 호흡기 감염의 원인: 중복검출의 임상적 의미)

  • Roh, Eui Jung;Chang, Young Pyo;Kim, Jae Kyung;Rheem, In Soo;Park, Kwi Sung;Chung, Eun Hee
    • Clinical and Experimental Pediatrics
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    • v.52 no.6
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    • pp.661-666
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    • 2009
  • Purpose : To determine the prevalence and clinical features of codetected respiratory etiological agents for acute respiratory infection in hospitalized children. Methods : Nasopharyngeal aspirates were obtained from hospitalized children with acute respiratory infection at Dankook University Hospital from September 2003 through June 2005. Immunofluorescent staining and culture were used for the detection of respiratory viruses (influenza virus [IFV] types A, B; parainfluenza virus [PIV] types 1, 2, 3; respiratory syncytial virus [RSV]; adenovirus [AdV]). Polymerase chain reaction (PCR) assays were used for Mycoplasma pneumoniae (MP) and Chlamydia trachomatis (CT) detection, and PCR and culture were performed for enterovirus detection. Acid-fast staining and culture were performed for tuberculosis detection. The demographic and clinical characteristics were reviewed retrospectively from the patients medical records. Results : Evidence of two or more microbes was found in 28 children: RSV was detected in 14, PIV 3 in 10, AdV in 10, MP in 8, PIV 2 in 8, CT in 4, and PIV 1 in 3. Codetected agents were found as follows: RSV+PIV 2, 6 patients; AdV+MP, 4 patients; AdV+PIV, 3 patients; RSV+MP, 3 patients; PIV 1+PIV 3, 3 patients. Distinct peaks of codetected agents were found in epidemics of MP and each respiratory virus. Conclusion : The codetected infectious agents were RSV, PIV, AdV, and MP, with distinct peaks found in epidemics of MP and each respiratory virus. Although advances in diagnostic methods have increased the prevalence of codetection, its clinical significance should be interpreted cautiously.

Detection of Vancomycin-Resistant Enterococci and Related Genes Using VITEK 2 System and Multiplex Real-time PCR Assay (VITEK 2 시스템과 Multiplex Real-time PCR을 이용한 반코마이신 내성 장알균(VRE)과 내성관련 유전자 검출)

  • Jeong, Min-Kyung;Yu, Young-Bin;Kim, Sang-Ha;Kim, Sunghyun;Kim, Young-Kwon
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.401-406
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    • 2017
  • In this study, using the VITEK 2 system, 74 samples (22.6%) out of 327 specimens were identified by the growth of Enterococcosel media (EV6 agar) supplemented with $6{\mu}g/mL$ of vancomycin. Enterococcus faecium was identified as 55 strains (74.3%), Enterococcus casseliflavus as 2 strains (2.7%), Enterococcus avium as 1 strain (1.4%), and Enterococcus gallinarum as 16 strains (21.6%). Among the 55 phenotypes of Enterococcus faecium, 42 (76.4%), 9 (16.4%), and 4 strains (7.3%) showed the vanA, vanB, and vanC phenotype, respectively. The 16 strains of Enterococcus gallinarum and 2 strains of Enterococcus casseliflavus showed the vanC phenotype and the 1 strain of Enterococcus avium had the vanB phenotype. The one strain of Enterococcus faecium propagated only in EV4 and was susceptible to both vancomycin and teicoplanin according to the antimicrobial susceptibility test using the VITEK 2 system. The vancomycin resistance phenotype gene was not detected by PCR. A total of 327 specimens were cultured in Enterococcosel broth supplemented with $6{\mu}g/mL$ of vancomycin (EV6 broth), and 120 strains (36.7%) were isolated. These 120 strains were subjected to vancomycin resistant genotyping by a multiplex real-time polymerase chain reaction and 51 strains (42.5%) showed vanA; 5 strains (4.2%) showed vanA and vanC; and 18 strains (15%) showed vanC. Vancomycin resistance genotypes were not detected in the remaining 46 strains (38.3%).