• 제목/요약/키워드: laboratory medical testing

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유전성 내분비 질환의 분자유전학적 진단 (Molecular Genetic Diagnosis of Genetic Endocrine Diseases)

  • 최진호;김구환;유한욱
    • Journal of Genetic Medicine
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    • 제7권1호
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    • pp.16-23
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    • 2010
  • 많은 내분비 질환이 유전적 요소를 갖고 있다. 단일 유전자 질환에서는 유전적 요인이 주요 원인이나 다인자성 질환에서는 환경과 생활습관 등이 함께 병인으로 작용한다. 유전성 내분비 질환의 분자유전학적 병인에 대한 이해에 대하여 최근 많은 발전이 있어 왔으며 분자유전학적 기술의 응용으로 질환에 대한 이해와 이를 이용한 진단 및 유전 상담에 도움이 되고 있다. 유전학적 검사로 특정 질환의 돌연변이를 증명하는 것은 진단이 모호한 경우에서 정확한 진단과 산전 진단, 보인자 검사에 적용될 수 있다. 그러나 유전자 검사만으로 임신 중절과 관련된 산전 진단에 이용하는 데에는 신중을 기해야 한다.

Clinical application of genome-wide single nucleotide polymorphism genotyping and karyomapping for preimplantation genetic testing of Charcot-Marie-Tooth disease

  • Kim, Min Jee;Park, Sun Ok;Hong, Ye Seul;Park, Eun A;Lee, Yu Bin;Choi, Byung-Ok;Lee, Kyung-Ah;Yu, Eun Jeong;Kang, Inn Soo
    • Journal of Genetic Medicine
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    • 제19권1호
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    • pp.7-13
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    • 2022
  • Purpose: Preimplantation genetic testing for monogenic disorders (PGT-M) has been successfully used to prevent couples with monogenic disorders from passing them on to their child. Charcot-Marie-Tooth Disease (CMT) is a genetic disorder characterized by progressive extremity muscle degeneration and loss of sensory function. For the first time in Korea, we report our experience of applying single nucleotide polymorphism genotyping and karyomapping for PGT-M of CMT disease. Materials and Methods: Prior to clinical PGT-M, preclinical tests were performed using genotypes of affected families to identify informative single-nucleotide polymorphisms associated with mutant alleles. We performed five cycles of in vitro fertilization PGT-M in four couples with CMT1A, CMT2A, and CMT2S in CHA Fertility Center, Seoul Station. Results: From July 2020 through August 2021, five cycles of PGT-M with karyomapping in four cases with CMT1 and CMT2 were analyzed retrospectively. A total of 17 blastocysts were biopsied and 15 embryos were successfully diagnosed (88.2%). Ten out of 15 embryos were diagnosed as unaffected (66.7%). Five cycles of PGT-M resulted in four transfer cycles, in which four embryos were transferred. Three clinical pregnancies were achieved (75%) and the prenatal diagnosis by amniocentesis for all three women confirmed PGT-M of karyomapping. One woman delivered a healthy baby uneventfully and two pregnancies are currently ongoing. Conclusion: This is the first report in Korea on the application of karyomapping in PGT-M for CMT patients. This study shows that karyomapping is an efficient, reliable and accurate diagnostic method for PGT-M in various types of CMT diseases.

유전자진단에 있어서 Multiplex Ligation Dependent Probe Amplification (MLPA)의 이론과 실제 (MLPA Applications in Genetic Testing)

  • 김구환;이범희;유한욱
    • Journal of Genetic Medicine
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    • 제6권2호
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    • pp.146-154
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    • 2009
  • Multiplex ligation dependent probe amplification (MLPA)은 탐침자를 표적지에 교잡시킨 후, ligation 시키고, 그 산물을 중합효소연쇄반응으로 증폭시킴으로써 표적지의 존재여부 또는 농도를 확인할 수 있는 방법으로, 그 원리가 소개된 이래로 여러 유전자들에 대한 거대결실 및 중복돌연변이에 대한 탐색에 이용되었다. 유전자진단은 질환에 관련된 유전자에 대한 돌연변이를 탐색함으로써 질환을 진단하는 방법으로, 단일유전자 결핍 질환에 대한 유전자진단은 주로 중합효소연쇄반응과 염기서열 분석 방법을 통한 점돌연변이의 탐색에 집중되어 있다. 거대결실 또는 중복돌연변이의 경우, 특히 이형접합자를 형성하게 되는 경우는 중합효소 연쇄반응을 통하여 결실 또는 중복돌연변이 여부의 확인이 힘들다. PCR 방법에 기초하여 유전자의 농도(gene dosage)를 알 수 있는 방법으로 MLPA 방법이 소개되면서 거대결실 또는중복돌연변이를 포함하고 있던 질병 관련돌연변이들의 규명이 한층 쉬워졌다. MLPA의 원리를 응용하여 단순한 유전자의 농도 측정뿐 아니라 유전자내의 메칠화양상의 차이를 확인하거나, 염색체의 배수체 이상 등 염색체이상의 돌연변이 규명과, 전체 유전자의 크기가 비교적 커서 거대결실 돌연변이를 많이 동반하는, 주로 우성유전의 암 관련 유전자 돌연변이의 규명에 유용하게 이용된다. MLPA는 상용적인 중합효소연쇄반응으로 확인할 수 없는 유전자의 농도를 효과적으로 규명할 수 있는 방법으로, 적은 양의 주형 DNA만을 사용하고, 한가지의 실험원리로 다양한 응용이 가능하며 high-throughput이 가능한 장점을 가지는 반면, 주형 DNA의 질에 결과의 의존도가 높고, 민족 또는 개인간의 차이를 보일 수 있는 표적 DNA 염기서열 내의 single nucleotide polymorphism (SNP) 등으로 인해 분석의 오류가 생길 수 있으며, 양적 차이를 규명하는 것이므로 수 차례의 대조군 검사가 함께 진행되어야 하는 단점이 있다. 여기서는 MLPA를 이용하여 질병유전자의 돌연변이를 밝힌 사례를 바탕으로 MLPA의 원리와 탐색할 수 있는 돌연변이의 종류, 그리고 이 방법의 장단점에 대해 고찰해 보고자 한다.

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A novel variant of PHEX in a Korean family with X-linked hypophosphatemic rickets

  • Kim, Sejin;Kim, Sungsoo;Kim, Namhee
    • Journal of Genetic Medicine
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    • 제19권1호
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    • pp.27-31
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    • 2022
  • X-linked dominant hypophosphatemic rickets are the most common form of familial hypophosphatemic rickets resulting from hypophosphatemia caused by renal phosphate wasting, which in turn is a result of loss-of-function mutations in PHEX. Herein, we report a 39-year-old female with short stature and skeletal deformities and 12-month-old asymptomatic daughter. The female has a history of multiple surgical treatments because of lower limb deformities. Her biochemical findings revealed low serum phosphorus levels with elevated serum alkaline phosphatase activity and normal serum calcium levels, suggesting presence of hypophosphatemic rickets. To identify the molecular causes, we used a multigene testing panel and found a mutation, c.667dup (p.Asp223GlyfsTer15), in PHEX gene. To the best of our knowledge, this is a novel mutation. A heterozygous form of the same variant was detected in daughter, who showed no typical symptoms such as bow legs, frontal bossing, or waddling gate, but presented early signs of impaired mineralization in both X-ray and biochemical findings. The daughter was initiated onto early medical treatment with oral phosphate supplementation and an active vitamin D analog. Because the daughter was genetically diagnosed based on a family history before the onset of symptoms, appropriate medical management was possible from early infancy.

Clinical Significance of Upregulation of mir-196a-5p in Gastric Cancer and Enriched KEGG Pathway Analysis of Target Genes

  • Li, Hai-Long;Xie, Shou-Pin;Yang, Ya-Li;Cheng, Ying-Xia;Zhang, Ying;Wang, Jing;Wang, Yong;Liu, Da-Long;Chen, Zhao-Feng;Zhou, Yong-Ning;Wu, Hong-Yan
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권5호
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    • pp.1781-1787
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    • 2015
  • Background: miRNAs are relatively recently discovered cancer biomarkers which have important implications for cancer early diagnosis, treatment and estimation of prognosis. Here we focussed on expression of mir-196a-5p in gastric cancer tissues and cell lines so as to analyse its significance for clinicopathologic characteristics and generate enriched KEGG pathways clustered by target genes for exploring its potential roles as a biomarker in gastric cancer. Materials and Methods: The expression of mir-196a-5p in poorly, moderate and well differentiated gastric cancer cell lines compared with GES-1 was detected by RT-qPCR, and the expression of mir-196a-5p in gastric cancer tissues comparing with adjacent non cancer tissues of 58 cases were also assessed by RT-qPCR. Subsequently, an analysis of clinical significance of mir-196a-5p in gastric cancer and enriched KEGG pathways was executed based on the miRWalk prediction database combined with bioinformatics tools DAVID 6.7 and Mirfocus 3.0. Results: RT-qPCR showed that mir-196a-5p was up-regulated in 6 poorly and moderate differentiated gastric cancer cell lines SGC-7901, MKN-45, MKN-28, MGC-803, BGC-823, HGC-27 compared with GES-1, but down-regulated in the highly differentiated gastric cancer cell line AGS. Clinical data indicated mir-196a-5p to beup-regulated in gastric cancer tissues (47/58). Overexpression of mir-196a-5p was associated with more extensive degree of lymph node metastasis and clinical stage (P < 0.05; x2 test). Enriched KEGG pathway analyses of predicted and validated targets in miRWalk combined with DAVID 6.7 and Mirfocus 3.0 showed that the targeted genes regulated by mir-196a-5p were involved in malignancy associated biology. Conclusions: Overexpression of mir-196a-5p is associated with lymph node metastasis and clinical stage, and enriched KEGG pathway analyses showed that targeted genes regulated by mir-196a-5p may contribute to tumorgenesis, suggesting roles as an oncogenic miRNA biomarker in gastric cancer.

Immortalization of Rabbit Corneal Fibroblasts by Overexpression of Simian Virus 40 Large T antigen

  • Cho, Seung-Ju;Park, Yuk-Pheel;Lim, Heon-Man;Kim, Jae-Chan;Yang, Eun-Kyung;Park, Jung-Keug;Yoon, Do-Young;Lee, Hee-Gu
    • 대한의생명과학회지
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    • 제10권2호
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    • pp.85-92
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    • 2004
  • Immortalization of primary corneal cells has influence on pharmacy, medical and biological fields. Especially, investigation of immortalization mechanism using viral oncoproteins is useful for medical treatments, and these cell lines will be useful materials for toxic test of medical supplies and cell biological experiments. Rabbit corneal fibroblasts in culture undergo a finite number of divisions before they reach a terminally non-proliferating state known as replicative senescence. Therefore, we attempted to induce immortalization of rabbit corneal fibroblasts with SV 40 large T antigen. As a result of experiment, expression of SV 40 large T antigen was confirmed, and expression of proteins related to cell cycle repressor was decreased in the transfection group compared with non-transfection group. According to the results of cell cycle phase distribution test, SV 40 large T antigen-transfected cells had obtained higher proliferation rate than primary cells. It was confirmed that during induction of immortalization, SV 40 large T antigen was not able to increase telomerase activity. In conclusion, we made a rabbit corneal fibroblast cell line with SV40 large T antigen. This cell line will be useful for further studies of mammalian fibroblast biology, particularly with regard to angiogenesis and malignant transformation. In addition, this cell line offers opportunity for testing potential therapeutics and can be used for toxicity tests of materials or cosmetics. In the future, our cell line can potentially be utilized in a wide range of biology related fields.

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Monitoring of Methicillin-resistant Staphylococcus aureus in Nasal Swabs Obtained from Dental Clinic Healthcare Providers and Medical Environment Nurses

  • Han, Seung-Ho;Song, In-Sook;Kim, Jong-Koan;Park, Jum-Gi;Park, Jang-Hwan;Lee, Myeong-Jae;Kim, Shin-Moo;Kim, Kang-Ju
    • International Journal of Oral Biology
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    • 제35권1호
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    • pp.7-12
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    • 2010
  • The aims of this study were to investigate the nosocomial infection route of methicillin-resistant Staphylococcus aureus (MRSA) and explore preventative methods for this pathogen that involve blocking its dispersion. We cultured MRSA from nasal cavity swabs collected between June and July 2008 that we obtained from eight dental healthcare providers, 32 nurses and the sputum specimens of two patients from our hospital. In addition, we used VITEK 2 equipment to measure drug sensitivity, and we further performed biochemical testing and pulse-field gel electrophoresis (PFGE) to isolate MRSA colonies. The incidence of these bacteria on the nasal swabs was 25.0% from dental clinic healthcare providers, 13.6% from the internal medicine ward nurses and 30.0% from intensive care unit nurses. Moreover, MRSA was detectable in sputum specimens of ward patients. The antimicrobial agents resistance and partial PFGE types of MRSA showed a similar pattern. We suggest from these analyses that nasal cavity infection by MRSA could occur by cross contamination between healthcare providers and patients which underscores the importance of stringent MRSA management practices.

Clinical utility of chromosomal microarray analysis to detect copy number variants: Experience in a single tertiary hospital

  • Park, Hee Sue;Kim, Aryun;Shin, Kyeong Seob;Son, Bo Ra
    • Journal of Genetic Medicine
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    • 제18권1호
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    • pp.31-37
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    • 2021
  • Purpose: To summarize the results of chromosomal microarray analysis (CMA) for copy number variants (CNVs) detection and clinical utility in a single tertiary hospital. Materials and Methods: We performed CMA in 46 patients over the course of two years. Detected CNVs were classified into five categories according to the American College of Medical Genetics and Genomics guidelines and correlated with clinical manifestations. Results: A total of 31 CNVs were detected in 19 patients, with a median CNV number per patient of two CNVs. Among these, 16 CNVs were classified as pathogenic (n=3) or likely pathogenic (LP) (n=11) or variant of uncertain significance (n=4). The 16p11.2 deletion and 16p13.11 deletion classified as LP were most often detected in 6.5% (3/46), retrospectively. CMA diagnostic yield was 24.3% (9/37 patients) for symptomatic patients. The CNVs results of the commercial newborn screening test using next generation sequencing platforms showed high concordance with CMA results. Conclusion: CMA seems useful as a first-tier test for developmental delay with or without congenital anomalies. However, the classification and interpretation of CMA still remained a challenge. Further research is needed for evidence-based interpretation.

Test turnaround Time for Complete Blood Cell Count using Delta and Panic Value Checks and the Q-flag Limit

  • Koo, Bon-Kyung;Ryu, Kwang-Hyun;Lim, Dae-Jin;Cho, Young-Kuk;Kim, Hee-Jin
    • 대한임상검사과학회지
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    • 제44권2호
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    • pp.66-74
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    • 2012
  • Test turnaround time (TAT) is the lead time from reception to reporting. In the complete blood cell count (CBC), 4 units of the XE-2100 (Sysmex Corp., Japan) processed around 80% of quantity, 1 unit of the LH-780 (Beckman-Coulter Incorp., USA) processed around 10% and 1 unit of ADVIA-2120 (Siemens AG, Munich, Germany) processed around 10%. We analyzed the change in the TAT for the CBC for over 7 years, from January of 2005 to December of 2011. The delta check made alterations of delta to WBC, hemoglobin, hematocrit, platelet and metamyelocyte, however, did not made them to band neutrophil, eosinophil, basophil and monocyte. The panic value check made alterations of panic value to hemoglobin, hematocrit, platelet and monocyte. In the criteria of currently slide review, LH-780 and ADVI-2120 analyzers prepared suspect flags of "Blast, Imm NE2, Immature granulocyte, Imm NE1, Left shift, Variant lymphocyte, Atypical lymphocyte, Platelet clumps and NRBC". The New slide review in the XE-2100 analyzer altered the preparations of a smear slide more than a "Platelet clumps flag(${\geq}200unit$), a single flag excluding the "Platelet clumps flag (${\geq}250unit$) and a multiple flag (${\geq}200unit$)". Also, below the 240 unit, medical technologists prepared manual slides selectively according to their evaluations. The automatic reporting rate was 33.4% without alterations, whereas it was 41.0% without alterations, and was thus improved by 7.6%. The slide review rate was 15.2% before using the Q-flag limit, whereas it was 12.1% for a reduce 3.1%. TAT was 45 minutes without the creation alterations of the delta and panic value checks, whereas it was 35 minutes after making alterations of the delta and panic value checks and thus was shortened by 10 minutes. We came to the conclusion that the establishment and operation of delta and panic value checks and slide review criteria suitable for laboratory environment can reduce unnecessary smear slides, re-checking, re-sampling, re-testing, telephone inquiries and concentrated workloads during specific times of the day.

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Distribution of Human Papilloma Virus Infections of Uterine Cervix among Women of Reproductive Age - a Cross Sectional Hospital-Based Study from North East India

  • Sarma, Usha;Mahanta, Jagadish;Borkakoty, Biswajyoti;Sarmah, Bidula
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권4호
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    • pp.1519-1523
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    • 2015
  • Infection of the uterine cervix by human papilloma viruses (HPV) may be associated with cervical pre-cancer and invasive cervical carcinoma if left untreated. With advance in molecular techniques, it has become easier to detect the resence of HPV DNA long before the appearance of any lesion. This study concerned cervical scrape samples of 310 married non-pregnant women attending a gynecology outpatient department for both Pap and PCR testing to detect HPV DNA. Nested PCR using primers for L1 consensus gene with My9/My11 and GP6+/GP5+followed by multiplex PCR were carried out to detect HPV 16 and HPV18. Result: HPV prevalence was 11.9% out of which 3.67% cases of negative for intra-epithelial lesion or malignancy (NILM) and in 71.1% (27/38) of atypical cervical smears were HPV positive. There was increasing trend of high-risk-HPV positivity (HR HPV 16 and 18), from 20% in benign cytology (NILM) to 42.9 % in LSIL, 71.41% in HSIL and 100% in SCC. There was highly significant association of HPV infection with cervical lesion ($x^2=144.0$, p<0.01) and also with type specific HPV prevalence ($x^2=7.761^*$, p<0.05).