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Real-time Reverse Transcription Polymerase Chain Reaction Using Total RNA Extracted from Nasopharyngeal Aspirates for Detection of Pneumococcal Carriage in Children (소아에서 폐렴구균 집락률 측정을 위해 비인두 흡인 물의 총 RNA를 이용한 실시간 중합효소 연쇄반응법)

  • Kim, Young Kwang;Lee, Kyoung Hoon;Yun, Ki Wook;Lee, Mi Kyung;Lim, In Seok
    • Pediatric Infection and Vaccine
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    • v.23 no.3
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    • pp.194-201
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    • 2016
  • Purpose: Monitoring pneumococcal carriage rates is important. We developed and evaluated the accuracy of a real-time reverse transcription polymerase chain reaction (RT-PCR) protocol for the detection of Streptococcus pneumoniae. Methods: In October 2014, 157 nasopharyngeal aspirates were collected from patients aged <18 years admitted to Chung-Ang University Hospital. We developed and evaluated a real-time PCR method for detecting S. pneumoniae by comparing culture findings with the results of the real-time PCR using genomic DNA (gDNA). Of 157 samples, 20 specimens were analyzed in order to compare the results of cultures, real-time PCR, and real-time RT-PCR. Results: The concordance rate between culture findings and the results of real-time PCR was 0.922 (P<0.01, Fisher exact test). The 133 culture-negative samples were confirmed to be negative for S. pneumoniae using real-time PCR. Of the remaining 24 culture-positive samples, 21 were identified as S. pneumonia -positive using real-time PCR. The results of real-time RT-PCR and real-time PCR from 20 specimens were consistent with culture findings for all S. pneumoniae -positive samples except one. Culture and real-time RT-PCR required 26.5 and 4.5 hours to perform, respectively. Conclusions: This study established a real-time RT-PCR method for the detection of pneumococcal carriage in the nasopharynx. Real-time RT-PCR is an accurate, convenient, and time-saving method; therefore, it may be useful for collecting epidemiologic data regarding pneumococcal carriage in children.

Utility of polymerase chain reaction(PCR) according to sampling time in CSF and stool specimens from patient with aseptic meningitis (무균성 뇌막염 환자에서 뇌척수액과 대변 채취 시점에 따른 PCR의 유용성)

  • Kim, Myo Jing;Lee, Hye Jin;Choi, Jung Mi;Jung, Soo Jin;Huh, Jae Won
    • Clinical and Experimental Pediatrics
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    • v.49 no.7
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    • pp.745-750
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    • 2006
  • Purpose : Enteroviruses are the most common cause of aseptic meningitis in patients of all ages. A definite diagnosis of enteroviral meningitis can be established by detection of virus directly in CSF specimens. But this is time-consuming and lacks sensitivity, so polymerase chain reaction(PCR) detecting of viral RNA in patient specimens such as CSF, stool has been demonstrated. But little is known about the influence of sampling time on the results of CSF PCR and stool PCR. We investigated diagnostic utility of PCR of CSF and stool according to sampling time after the onset of symptoms. Methods : PCR results were analyzed according to sampling time for 42 patients diagnosed aseptic meningits in our hospital from $11^{th}$ January to $30^{th}$ August, 2005. Results : The diagnostic yield of the test was higher of CSF specimens obtained ${\leq_-}2$ days after clinical onset(positive PCR results 9/18, 50 percent), compared with CSF collected >2 days after onset(positive PCR results 1/24, 4.2 percent)(P=0.001). Instead, positive PCR results of fecal specimens maintained highly(average 90.5 percent), 10 cases had also positive PCR results even 5-6 days after onset. 10 cases of CSF specimens had positive enterovirus PCR results containing coxsackievirus B5 (n=6), coxsackievirus B3(n=3). 38 cases of stool specimens had positive enterovirus PCR results containing echovirus 18(n=7), echovirus 9(n=3), coxsackievirus B5(n=8), coxsackievirus B3(n=3). 6 cases(coxackie B5) had positive CSF PCR and stool PCR, both. Conclusion : Stool PCR was clinically sensitive for detecting enterovirus during enteroviral meningits and could give a presumptive diagnosis throughout the disease course. A definite diagnosis was obtained by CSF PCR, but its utility was clearly lower for samples obtained >2 days after clinical onset. Therefore, it is recommended that, in addition to performance of CSF PCR, fecal samples obtained from patients with suspected enteroviral meningitis should be tested by PCR, especially when the duration of symptoms is >2 days.

Report on the Effects Lipemic Specimen in Anti-ds DNA Antibody Test (Anti-ds DNA 항체 검사 시 Lipemic 검체의 영향에 관한 보고)

  • Cheon, Jun Hong;Kim, Whe Jung;Kim, Sung Ho;Moon, Hyoung Ho;Yoo, Seon Hee
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.153-157
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    • 2014
  • Purpose: SLE (systemic lupus erythematosus) is an inflammatory autoimmune disease, characterized by various autoantibody. The detection of Anti double-stranded DNA (Anti-ds DNA) is important in the diagnostics of SLE, and include the American College of Rheumatology (ACR) diagnostic criteria for SLE. Also SLE disease activity and correlativity with the level Anti-ds DNA antibody have been reported and Anti-ds DNA antibody quantitative test is very useful for tracing before and after SLE treatment. When These Anti-ds DNA antibody test (Farr assay: $^{125}I$ labeled ds-DNA and bound Anti-ds DNA antibodies complex in serum is precipitated by ammonium sulfate and used to centrifugation, measured it) inhaled supernatant after centrifugation, a lipemic specimen does not facilitate the formation of precipitate and also occurs situation was inhaled with precipitate. To solve these problems, The Influence of the degree of lipemic specimen was evaluated. Materials and Methods: September 2012 to February 2013, We selected lipemic samples (n=81) of specimen commissioned by Anti-ds DNA antibody test. Lipemic samples were done pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) used a micro-centrifuge (Eppendorf Model 5415D). At the same time lipemic specimen and pre-treatment samples were performed Anti-ds DNA antibody test (Anti-ds DNA kit, Trinity Biotech, Ireland). Statistical analysis were analyzed Pearson's correlation coefficients and regression and paired t-test, and Difference (%). Results: Experimental group 1 (Lipemic Specimen Anti-ds DNA Ab concentration ${\leq}7IU/mL$) at y=0.368X+4.732, $R^2=0.023$, Pearson's correlation coefficient was 0.154, paired t-test (P=0.003), Difference (%) mean 65.7 and showed a statistically significant difference. Experimental group 2 (Lipemic Specimen Anti-ds DNA Ab concentration ${\geq}8IU/mL$) at y=0.983X+0.298, $R^2=0.994$, Pearson's correlation coefficient showed 0.997, paired t-test (P=0.181), Difference (%) mean -5.53 made no statistically significant difference. Conclusion: Lipemic sample of low Anti-ds DNA Ab concentration (2.5-7 IU/mL) and the result is obtained pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) were made a significant difference statistically. Anti-ds DNA is one of the primary auto-antibodies present in patients with SLE, and remain an important diagnostic test for SLE. Therefore, we recommend preprocessing (high-speed centrifugation: 14,000 rpm 5 mins) in order to exclude the influence of lipemic specimen.

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Analysis of Clostridium Difficile Toxin Value in Diarrhea Patients (설사증 환자에서 Clostridium Difficile Toxin Value 분석)

  • Kwon, Se-Young;Yoon, In-Sook
    • The Journal of the Korea Contents Association
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    • v.10 no.5
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    • pp.259-266
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    • 2010
  • Clostridium difficile-associated disease (CDAD) is an important nosocomial infectious diarrhea and is associated with antibiotic use. Recently, incidence of C. difficile has been increasing in hospitals. A total of 1,329 stool specimens were examined from January, 2005 to December, 2008. This study analyzed the incidence and clinical characteristics of C. difficile infections on them. Out of 1,329 stool specimens, 283 specimens showed toxin A/B positive, using EIA. The positive rate was 21.2%; with the highest incidence among and above the 70years old. On endoscopy, psedo membranous colitis was found in 57.7%, and 19.5% of patients were normal. Pathologic finding showed PMC in 26.8% of patients, AAC in 52.2%. C. difficile was associated with PMC, however, endoscopic and pathologic findings tests showed normal to PMC.

Clinical Specimen Printing System using RFID (RFID를 이용한 검체 프린팅 시스템)

  • Kim, Yong-Phil;Choi, Kwang-Il;Jung, Hoe-Kyung
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.18 no.2
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    • pp.351-356
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    • 2014
  • Although the demand for histopathological examinations has been increasing, medical accidents in management of specimens also have been increasing because most of the examinations are processed manually which can cause careless handing, confusing information and mismatching during the procedure. In the future, histopatological examination will be used frequently for handing incurable diseases and verification of new drug. Thus, efficient and error-free management system for handling personalized medical history and test results is infallibly necessary. In this paper, I have proposed an integrated printing system for informatization of histopathological examination that support the u-Healthcare environment based on RFID in near future. The proposed system supports systematization of whole examination process and information of pathological samples. This system will contribute to reduction of costs, improvement of operational efficiency, and mostly fundamental prevention of medical accidents.

Expression of Micro RNA in Paraffin Embedded Tissue of Multiple Myeloma (다발성골수종 환자의 파라핀포매조직에서 MicroRNA 발현)

  • Choi, Woo Soon;Kwon, Kye Chul
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.292-297
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    • 2015
  • Research of thyroid cancer, liver cancer, and lung cancer has been reported in Korea. However microRNA research of multiple myeloma has never been reported. Hence we intended to confirm whether microRNA can be utilized as a diagnostic marker to patients of multiple myeloma. We also intended to evaluate whether microRNA can be detected in paraffin-embedded tissue (FFPE). This research was conducted targeting 8 samples from patients of multiple myeloma who do not have any other diseases, and 2 control samples. From January 2010 to July 2012, we selected miR-15a, miR-16, miR-21, miR-181a and miR-221 as microRNA target genes. It was decided that for a sample to be significant, the results should show values more than 1.5 or less than -1.5. Our findings of fold change were highly significant in miR-15a with a value of 37.5% (3/8). From these studies, we learned that miR-15a is useful with westerners. miR-221, on the other hand, shows conflicts with westerners, so more research will be needed in this area. In addition, it was confirmed that microRNA can be detected in paraffin embedded tissue (FFPE).

The Prevention of Carryover in the Automated System With Fixed Tips (자동 분주기 Fixed Tip의 Carryover 방지를 위한 개선 방안)

  • Hwang, Bo-Ra;Au, Doo-Hee;Bae, Jin-Su;Park, Jong-O;Kim, Ji-Young;Seok, Jae-Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.13 no.3
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    • pp.181-184
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    • 2009
  • Purpose: Automated system that immunoassay examination are used widely. However, sample to sample carryover can cause that the next patient sample is false positive. Materials and Methods: We test HBs Ag, HBs Ab, HBc Ab(IgG) with Automated pipetting system (Tecan). It was placed with very high concentrations followed by saline solution. During this experiment, The fixed tip of Automated system wash With 0.25 moL/L NaOH. The Measurement results of saline solution confirm the occurrence of carryover. Results: Results of saline solution with 0.25 moL/L NaOH cleaning process was measured 100% negative, And results of patient serum with 0.25 moL/L NaOH cleaning process is similar reported results. Conclusion: As Results, 0.25 moL/L NaOH cleaning process was avoid carryover in experiment, And we know results of the hepatitis test did not affected by this solution we recommend 0.25 moL/L NaOH cleaning process as the Prevention of carryover in the automated system with fixed Tips.

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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.

Fine Needle Aspiration Cytology vs. Core Needle Biopsy for Thyroid Nodules: A Prospective, Experimental Study Using Surgical Specimen (갑상선 결절에 대한 세침 흡인 세포 검사와 중심부 바늘 생검의 비교: 수술 검체를 기반으로 한 전향적, 실험적 비교 연구)

  • Hyuk Kwon;Jandee Lee;Soon Won Hong;Hyeong Ju Kwon;Jin Young Kwak;Jung Hyun Yoon
    • Journal of the Korean Society of Radiology
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    • v.83 no.3
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    • pp.645-657
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    • 2022
  • Purpose To evaluate and compare the diagnostic outcomes of ultrasonography (US)-guided fine needle aspiration (FNA) and core needle biopsy (CNB) performed on the same thyroid nodule using a surgical specimen for direct comparison. Materials and Methods We included 89 thyroid nodules from 88 patients from February 2015 to January 2016. The inclusion criterion was thyroid nodules measuring ≥ 20 mm (mean size: 40.0 ± 15.3 mm). Immediately after surgical resection, FNA and subsequent CNB were performed on the surgical specimen under US guidance. FNA and CNB cytopathologic results on the specimen were compared with the surgical diagnosis. Results Among the 89 nodules, 30 were malignant and 59 were benign. Significantly higher inconclusive rates were seen in FNA for malignant than benign nodules (80.0% vs. 39.0%, p < 0.001). For CNB, conclusive and inconclusive rates did not differ between benign and malignant nodules (p = 0.796). Higher inconclusive rates were seen for FNA among cancers regardless of US features, and in the subgroup of size ≥ 40 mm (62.5% vs. 22.9%, p = 0.028). Eleven cancers were diagnosed with CNB (36.7%, 11/30), while none was diagnosed using FNA. Conclusion In this experimental study using surgical specimens, CNB showed a potential to provide improved diagnostic sensitivity for thyroid cancer, especially when a conclusive diagnosis is limited with FNA.

US Features of the Parathyroid Glands: An Intraoperative Surgical Specimen Study (부갑상선의 초음파 소견: 수술 중 수술 검체 연구)

  • Wooyul Paik;Jong Cheol Lee;Byeong-Joo Noh;Dong Gyu Na
    • Journal of the Korean Society of Radiology
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    • v.84 no.3
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    • pp.596-605
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    • 2023
  • Purpose This study aimed to evaluate the US features of the parathyroid glands (PTGs) using surgical specimens of normal PTGs obtained during thyroid surgery. Materials and Methods This study included 34 normal PTGs from 17 consecutive patients who underwent thyroid surgery between December 2020 and March 2021. All normal PTGs were histologically confirmed by intraoperative frozen-section biopsy for autotransplantation. Surgically resected parathyroid specimens were scanned in sterile normal saline using high-resolution US prior to autotransplantation. The US features of echogenicity (hyperechogenicity or hypoechogenicity), echotexture (homogeneous or heterogeneous), size, and shape (ovoid or round) were retrospectively evaluated. The echogenicity of the three PTGs was compared with that of the thyroid parenchyma of the resected thyroid specimens in two patients. Results All PTGs showed hyperechogenicity similar to that of gauze soaked in normal saline. Homogeneous hyperechogenicity was observed in 32/34 (94.1%) patients, and the echogenicity of the three PTGs was hyperechoic compared with that of the thyroid parenchyma. The long diameter of the PTGs ranged from 5.1 mm to 9.8 mm (mean, 7.1 mm) and the shape of the PTGs was ovoid in 33/34 (97.1%) patients. Conclusion The echogenicity of normal PTG specimens was consistently hyperechoic, and the small ovoid homogeneously hyperechoic structure was a characteristic US feature of the PTGs.