• Title/Summary/Keyword: Blood PCR

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Utilization of Genetic Polymorphisms in Cattle (소에서의 유전적 다형의 이용)

  • 신형두
    • Journal of Embryo Transfer
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    • v.10 no.1
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    • pp.23-31
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    • 1995
  • Several kinds of analytic methods for genetic polymorphism in cattle, including bovine blood typing, PCR-RFLP, BoLA and microsatellite typing were described. A few respect to consider for choosing method for actual application of genetic polymorphism were emphasized. The probability of relationship between characteristics and gene concerned, repetibility and easiness of methods applied and the possibility of clarification for segregation pattern should be deliberated.

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Detection of Lymphotropic Herpesviruses by Multiplex Polymerase Chain Reaction

  • Park, Sang-Tae;Kim, Seung-Han;Lee, Dong-Gun;Park, Jung-Hyun;Shin, Wan-Shik;Kim, Tai-Gyu;Paik, Soon-Young;Kim, Chun-Choo
    • Journal of Microbiology
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    • v.39 no.3
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    • pp.226-228
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    • 2001
  • Human lymphotropic herpesvirus is known to be a major pathogen associated with various diseases in bone marrow transplantation (BMT) recipients. A multiplex nested-polymerase chain reaction (PCR) method was developed for the simultaneous detection of human lymphotropic herpesviruses, including Ebstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 variants A and B (HHV6-A, HHV6-B). To demonstrate the usefulness of multiplex PCR for the analysis of clinical samples, peripheral blood mononuclear cells and serum from BMT recipients were analysed. The results skewed that a clear detection could be made between EBV, HCMV and HHV-6. This multiplex PCR assay is an efficient and cost-effective approach to the analysis of large numbers of samples to determine the epidemiological importance of EBV HCMV and HHV-6.

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Detection of HBV Resistance to Lamivudine in Patients with Chronic Hepatitis B Using Zip Nucleic Acid Probes in Kerman, Southeast of Iran

  • Afshar, Reza Malekpour;Mollaie, Hamid Reza
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3657-3661
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    • 2012
  • HBV infection is contagious and may be transmitted vertically or horizontally by blood products and body secretions. Over 50% of Iranian carriers have contracted the infection prenatally, making this the most likely route of transmission of HBV in Iran. This study assesses the resistance to Lamivudine in patients with chronic hepatitis B infection using a new ZNA probe Real Time PCR method. To evaluate the effectiveness of Lamivudine therapy for chronic hepatitis B infection, a study was conducted on 70 patients (63 men and 7women), who had received the drug first line. All patients were tested for the presence of HBsAg and HBeAg, the serum ALT level and the HBV DNA load before and after treatment. In all samples resistance to Lamivudine was tested with the ZNA Probe. Our results showed that ZNA Probe Real Time PCR method could detect wild type,YMDD, and its mutants, tyrosine-isoleucine-aspartate-aspartate and tyrosine-valine-aspartate-Aspartate. Among an estimated seventy patients with chronic hepatitis B infection, 18 (25.7%) were resistant to lamivudine. Only one patient was negative for presence of HBS-Ag (5.6%) and two patients were negative for HBe-Ag (11.1%). Real-time PCR with Zip nucleic acid probes is a sensitive, specific and rapid detection method for mutations in the YMDD motif, which will be essential for monitoring patients undergoing Lamivudine antiviral therapy.

Expression of Endometriosis Related Genes in the Shed Endometrial Tissues from Menstrual Blood (생리혈에 존재하는 자궁내막조직에서 자궁내막증 관련 유전자의 발현 양상)

  • Park, Chan-Woo;Jun, Jin-Hyun;Koong, Mi-Kyoung;Song, In-Ok
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.4
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    • pp.275-283
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    • 2007
  • Objective: This study was performed to investigate the expressions of endometriosis related genes in shed endometrial tissues from menstrual blood of patients with or without endometriosis. Methods: The shed endometrial tissues were collected on 2$^{nd}$ or 3$^{rd}$ day of menstrual cycle with Wallace catheter in patients with endometriosis (n=16) and without endometriosis (n=26). The mRNA expressions of twelve kinds of endometriosis related genes were compared between two groups using semi-quantitative RT-PCR. Results: The collected shed endometrium was confirmed by histological observation. Expressions of telomerase, c-kit and aromatase mRNA were not detected by RT-PCR in shed endometrial tissues. The mRNA expressions of apoptosis related genes (fas, fas ligand, bcl-2, bax), stem cell factor, estrogen receptor-$\alpha$/$\alpha$, endometriosis protein-I and secretory leukocyte protease inhibitor gene were similar between shed endometrial tissues with endometriosis and without endometriosis. Conclusion: We could not find the difference of mRNA expressions of tested endometriosis related genes between shed endometrial tissues with or without endometriosis by semi-quantitative RT-PCR analysis. It may be related to the dynamical changes of gene expressions in the endometrium with menstrual cycle.

Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing (차세대염기서열분석법을 이용한 HLA-A, -B 그리고 -DR 형별 분석법 개발)

  • Seo, Dong Hee;Lee, Jeong Min;Park, Mi Ok;Lee, Hyun Ju;Moon, Seo Yoon;Oh, Mijin;Kim, So Young;Lee, Sang-Heon;Hyeong, Ki-Eun;Hu, Hae-Jin;Cho, Dae-Yeon
    • The Korean Journal of Blood Transfusion
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    • v.29 no.3
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    • pp.310-319
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    • 2018
  • Background: Research on next-generation sequencing (NGS)-based HLA typing is active. To resolve the phase ambiguity and long turn-around-time of conventional high resolution HLA typing, this study developed a NGS-based high resolution HLA typing method that can handle large-scale samples within an efficient testing time. Methods: For HLA NGS, the condition of nucleic acid extraction, library construction, PCR mechanism, and HLA typing with bioinformatics were developed. To confirm the accuracy of the NGS-based HLA typing method, the results of 192 samples HLA typed by SSOP and 28 samples typed by SBT compared to NGS-based HLA-A, -B and -DR typing. Results: DNA library construction through two-step PCR, NGS sequencing with MiSeq (Illumina Inc., San Diego, USA), and the data analysis platform were established. NGS-based HLA typing results were compatible with known HLA types from 220 blood samples. Conclusion: The NSG-based HLA typing method could handle large volume samples with high-throughput. Therefore, it would be useful for HLA typing of bone marrow donation volunteers.

Dietary Evaluation and Protein Catabolic Rate in Maintenance Hemodialysis Patients (혈액투석환자의 식이조사 및 Protein Catabolic Rate에 관한 연구)

  • 장유경
    • Journal of Nutrition and Health
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    • v.25 no.3
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    • pp.256-263
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    • 1992
  • As various metabolic alterations develope in uremic patients. their diets need to be restricted, Furthermore medical complications with accompanying anorexia result in further complications and decrease in body strength. To assess the nutritional status of hemodialyzed patients we performed evaluation for dietary intake and protein catabolic rate(PCR) For 24 clinically stable male patients undergoing maintenance hemodialysis dietary intake was estimated by 3-day food record method and PCR was calculated with blood urea nitrogen at pre and post hemodialysis. The results were as follows : 1) Average daily energy and protein intake were 26.7$\pm$5.1kcal/kg of body weight. 0.95$\pm$0.19 g/kg of body weight respectively. 2) Protein catabolic rate calculated from interdialysis blood urea nitrogen levels was 1.00$\pm$0.20g/kg of body weight. Protein catabolic rate was correlated with the amount of Protein intake(r=0.44 p<0.05) 3) Relative body weight(RBW) of the subjects was smaller than that of healthy man without hemodialysis. Calorie and protein intake and protein catabolic rate were significantly different (p<0.05) between patients with lower RBW(<90% of ideal body weight) and those with normal RBW(90~110% of ideal body weight) and those with normal RBW(90~110% of iedal body weight) 4) The duration of hemodialysis did not have a significant effect on the nutritional status of the subjects.

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Beagle dogs parentage testing by using 22 ISAG microsatellite markers

  • Ji, Hye-jung;Kim, Eun-hee;Lee, Kyoung-kap;Kang, Tae-young;Lee, Joo-myoung;Shin, Hyoung-doo;Kim, Lyoung-hyo;Yun, Young-min
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.457-460
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    • 2007
  • The objective of the study was to establish routine parentage testing system in Beagle dogs using 22 ISAG (International Society for Animal Genetics) canine microsatellite markers (2005). Blood collections were obtained from a mother dog, 4 candidate father dogs and 3 offspring (n = 8). Genomic DNA samples were extracted from 8 Beagle dogs blood for PCR analysis. PCR products for the allele were analyzed by ABI 3130 DNA Sequencer and GeneScan (Ver 3.0) analysis and Genotyper (Ver. 2.1) software. The genetic relationship of mother and 3 offspring as well as one father dog among 4 candidate father dogs was confirmed by microsatellite allele analysis. The results of locus for amelogenin, which was designed for sexing, were matching with real gender among 8 Beagle dogs (female; 217/217 homozygosity, male; 179/217 heterozygosity). Twenty two ISAG microsatellite markers are useful the parentage test of Beagle dogs. In addition, amelogenin is an applicable marker to detecting real sex in dogs.

Blood Glucose Lowering Activity and Mechanism of Sangbackpitang (SBPT) in db/db Mouse (db/db 마우스에서 상백피탕의 혈당강하 활성 및 기전연구)

  • 이성현;안세영;두호경;정성현
    • YAKHAK HOEJI
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    • v.43 no.6
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    • pp.818-826
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    • 1999
  • Antidiabetic activity and mechanism of Sangbackpitang (SBPT) was examined in db/db mice, which is a spontaneously hyperglycemic, hyperinsulinemic and obese animal model. SBPT and acarbose were administered orally for 4 weeks. Fasting and non-fasting serum glucose, glycated hemoglobin and triglyceride were all reduced when compared between db/db control group and SBPT treated group. At 12th week after birth, SBPT increased an insulin secretion although statistic significance was not seen. Total activities of sucrase, maltase and lactase in SBPT treated group were all decreased when compared to db/db control. On the other hand, sucrase and maltase activities in acarbose treated groups were increased. Effect of SBPT on mRNA expression of glucose transporter(GLUT-4) was also examined. Quantitation of glucose transporter was performed by RT-PCR and in vitro transcription with co-amplification of rat-action gene as an internal standard. Muscular GLUT-4 mRNA expression in SBPT treated group was increased significantly. These results may suggest that SBPT lowered blood glucose ascribing to inhibition of glycosidase-catalyzed reaction and upregulation of muscular GLUT-4 mRNA expression.

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Imported human babesiosis in the Republic of Korea, 2019: two case reports

  • Hyun Jung Kim;Min Jae Kim;Hyun-Il Shin;Jung-Won Ju;Hee-Il Lee
    • Parasites, Hosts and Diseases
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    • v.61 no.1
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    • pp.72-77
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    • 2023
  • Human babesiosis is a tick-borne disease induced by the genus Babesia and has been significantly reported in the Republic of Korea. This report shows the cases of 2 patients with human babesiosis who traveled to the USA in 2019. The 2 patients experienced fever and had travel histories to babesiosis-endemic regions. The diagnoses of both cases were verified by the identification of Babesia-infected red blood cells on blood smears. One patient was found to be infected with Babesia microti using polymerase chain reaction (PCR) for 18S rRNA, which discovered the phylogenetic link to the B. microti strain endemic in the USA. The 2 patients recovered from fever with subsequent hemoparasite clearance. Babesiosis could be diagnosed in anyone with histories of travel to babesiosis-endemic countries and tick bites. Furthermore, Babesia-specific PCR is required for determining geno-and phenotypic characteristics.