• Title/Summary/Keyword: Clinical samples

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Evaluating the results of the Momguard noninvasive prenatal test

  • Hu, Hae-Jin;Kwon, Young-Jun;Oh, Mijin;Kim, Jihun;Cho, Dae-Yeon;Seo, Dong-Hee
    • Journal of Genetic Medicine
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    • v.12 no.2
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    • pp.96-99
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    • 2015
  • Purpose: To evaluate the performance of the Momguard noninvasive prenatal test by tracing the 'screen positive' results based on preliminary samples from Korean cohorts. Materials and Methods: This preliminary study is based on data collected by the LabGenomics Clinical Laboratory (Seongnam, Korea) with informed consent. Only pregnant women who underwent both the Momguard test and karyotyping were included in this study. Momguard test results were compared with those of the karyotyping analysis. Results: Among the 38 cases with 'screen positive' results by Momguard, 30 cases also had karyotyping results available. In three trisomy (T) 18 and three T13 cases, the Momguard results were concordant with the karyotyping results. For the T21 cases, except for one case belonging to the mid-risk zone, Momguard results from 23 out of 24 cases matched the karyotyping results. Conclusion: Momguard is a highly reliable screening tool for detecting T13, T18, and T21 cases in independent Korean cohort samples.

Countermeasures of Specimens Showing below Reference Value in APTT Result

  • Choi, Hyun Oh;Kim, Seung Gu;Park, Sang Hee;Lee, Jae Ki;Koo, Bon Kyung;Park, Chang Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.3
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    • pp.87-90
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    • 2013
  • The activated partial thromboplastin time (APTT) is used primarily to evaluate coagulation abnormalities in the intrinsic pathway. The proper specimen is very important factor for precise results of blood coagulation analysis. The objective of this study was to get to the conclusion of whether to analyze again and to collect blood sample over again when APTT result shows below the reference value. We evaluated 126 samples showing a value below 20.0 sec at ATPT result, which consist of 48 males and 78 females candidates during night duty from March 2012 to December 2012. Average comparisons of APTT result between first and retested analysis among study subjects were significantly different in male samples. APTT results comparison of recollected subjects among clotted samples were also significantly different with both sexes (p<0.000). We suggest that we should carefully check the samples to get accurate results and collect samples again in case of only obtaining improper samples; even though the APTT result show below reference value.

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Detection of Virulence-Associated Genes in Clinical Isolates of Bacillus anthracis by Multiplex PCR and DNA Probes

  • Kumar, Sanjay;Tuteja, Urmil
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1475-1481
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    • 2009
  • Anthrax is a zoonotic disease caused by Bacillus anthracis, and well recognized as a potential agent for bioterrorism. B. anthracis can be identified by detecting the virulence factors genes located on two plasmids, pXO1 and pXO2. The aim of the present study was to determine the presence of virulence genes in 27 isolates of B. anthracis isolated from clinical and environmental samples. For this purpose, multiplex PCR and DNA probes were designed to detect protective antigen (pag), edema factor (cya), lethal factor (lef), and capsule (cap) genes. Our results indicated that all the isolates contained all the above virulence genes, suggesting that the isolates were virulent. To the best our knowledge, this is the first study about the determination of virulence marker genes in clinical and environmental isolates of B. anthracis using multiplex PCR and DNA probes in India. We suggest that the above methods can be useful in specific identification of virulent B. anthracis in clinical and environmental samples.

Molecular detection and genotype analysis of Kudoa septempunctata from food poisoning outbreaks in Korea

  • Gyung-Hye Sung;In-Ji Park;Hee-Soo Koo;Eun-Hee Park;Mi-Ok Lee
    • Parasites, Hosts and Diseases
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    • v.61 no.1
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    • pp.15-23
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    • 2023
  • Concerns about foodborne illnesses caused by Kudoa septempunctata are steadily growing, but reports of K. septempunctata in clinical and food specimens related to food poisoning in Korea are limited. This study aimed to genetically identify K. septempunctata in patients with acute diarrhea and in clinical and food samples related to food poisoning caused by sashimi consumption. Both real-time and nested polymerase chain reaction assays were performed to detect K. septempunctata 18S and 28S rDNA genes in the stools of 348 patients with acute diarrhea, 11 samples (6 stool and 5 rectal swab samples) from patients with food poisoning, and 2 raw Paralichthys olivaceus samples collected from a restaurant where a food poisoning incident occurred. K. septempunctata was identified in 5 clinical specimens (4 stools and 1 rectal swab) and 1 P. olivaceus sashimi sample. All detected K. septempunctata were of genotype ST3. This is the first study to identify K. septempunctata in both patients and food samples with epidemiological relevance in Korea, providing evidence that it is a pathogen that causes food poisoning. Also, this is the first study to confirm the presence of K. septempunctata genes in rectal swabs. Despite continuing suspected occurrences of Kudoa foodborne outbreaks, the rate of identification of K. septempunctata is very low. One reason for this is the limitation in obtaining stool and vomit samples for the diagnosis of Kudoa infection. We strongly suggest the inclusion of rectal swabs among the diagnostic specimens for Kudoa food poisoning.

Production of Plasma Leptin and Expression of Interferon-γ Inducible Protein-10 (IP-10), Monokine Induced by Interferon-γ (Mig) and Interleukin-8 (IL-8) mRNA in Kawasaki Disease

  • Lee, Young-Hwan;Kim, Won-Duk;Kim, Hee-Sun
    • IMMUNE NETWORK
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    • v.2 no.4
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    • pp.202-207
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    • 2002
  • Background: Kawasaki disease is an acute febrile illness with systemic vasculitis which primarily affects children, We examined the production of leptin in plasma and gene expressions of CXC chemokines in peripheral blood mononuclear cells from patients with Kawasaki disease. Methods: Consecutive 39 samples from 13 patients according to the different clinical stages (acute, subacute, convalescent) of Kawasaki disease were collected. The plasma leptin levels according to clinical stages of Kawasaki disease were examined by ELISA and the expression of IP-10, Mig and IL-8 mRNAs in 39 samples (13 samples of each stage) from 13 cases were examined by RT-PCR. Results: There were not significant changes of plasma leptin levels according to the clinical stages of Kawasaki disease. The mean values of plasma leptin concentrations during each of the stages (n=13, p>0.05, pg/ml) were $335.8{\pm}549.0$ in acute, $358{\pm}347.6$ in subacute, and $443.6{\pm}645.9$ in convalescent stage. The mRNAs of IP-10, Mig, and IL-8 were expressed in 13/13 (100%), 2/13 (15%), 9/13 (69%) during acute stage, 13/13 (100%), 6/13 (46%), 13/13 (100%) during subacute stage, and 13/13 (100%), 4/13 (31%), 10/13 (77%) during the convalescent stage, respectively. In three patients, the production of leptin and expression of IP-10 mRNA were dramatically decreased according to the process of the clinical stages. In five patients with prominent cervical lymphadenopathy, the expression of IL-8 mRNA during the subacute stage was more elevated than the acute and convalescent stages. Conclusion: This data suggests that the production of leptin and the gene expressions of IP-10, Mig and IL-8 seem to have no significant correlation to the clinical stages of Kawasaki disease. However, expression patterns of IP-10, Mig and IL-8 mRNA may be related to the specific clinical manifestations, and the expression of IP-10 may also be correlated to leptin levels with pericardial involvement.

In vitro Study on the Antimicrobial Activity of Human Tears with Respect to Age

  • Zahoor, Muhammad;Bahadar, Haji;Ayaz, Muhammad;Khan, Ajmal;Shah, Muhammad Jalat
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.2
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    • pp.93-99
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    • 2018
  • Lysozyme is present in tears and has the ability to inhibit bacterial growth. In addition, it acts as a physiological scavenger for harmful substances. In the present study, sixteen tear samples from people of different ages were evaluated for their antibacterial spectrum against selected bacterial strains (Escherichia coli, Shigella sonnei, Staphylococcus aureus, Salmonella enterica Typhi). A radial diffusion assay was used to evaluate the antibacterial potential of tear samples. To correlate the antibacterial activities of these tear samples, the concentration of lysozyme in the tear samples was also determined. Ampicillin was used as a standard drug. The zone of inhibition (mm) was used to measure the antibacterial property of the tears. All samples showed good antibacterial activities. The tear samples of children showed antibacterial activities in the range of 4.40~5.00 mm inhibition zones against the selected bacterial strains. The tear samples from the young and adults showed good antibacterial potential with a zone of inhibition in the range of 3.20~4.00 and 4.00~5.50 mm, respectively. The tear samples from the old age group showed inhibition zones from 1.50~5 mm. The adult tear samples showed the maximum inhibition against the selected bacterial strains among all groups. The lysozyme concentration was 1.7 mg/mL, 1.95 mg/mL, 2.13 mg/mL, and 1.76 mg/mL for children, young, adults, and elderly, respectively. In conclusion, the tears from adults have the high inhibition potential. In addition, this data also showed that the lysozyme contents in the tear sample increased with age until 40~42 years.

Bacterial Contamination of Dental Unit Water Systems in a Student Clinical Simulation Laboratory of College of Dentistry (치과대학 임상 시뮬레이션 실습실 치과용 유니트 수계의 세균 오염도 조사)

  • Yoon, Hye Young;Lee, Si Young
    • Journal of dental hygiene science
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    • v.15 no.2
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    • pp.232-237
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    • 2015
  • The water supplied from dental unit water systems (DUWS) in dentistry may be heavily contaminated with bacteria and thus may be a potential source of infection for both practice staff and patients. The aim of this study was to evaluate the level of heterotrophic bacteria and to confirm the presence of opportunistic pathogens from DUWS in student clinical simulation laboratory of college of dentistry. Water samples were collected from 36 ultrasonic scalers in student clinical simulation laboratory. The levels of heterotrophic bacteria in water samples were quantified by counting colony forming units (CFUs) on R2A agar media. In addition, opportunistic pathogens were detected by using the polymerase chain reaction (PCR) method. The mean CFUs were 16,095 CFU/ml for water samples and all of water samples exceeded current American Dental Association recommendations of 200 CFU/ml. Pseudomonas species and non-tuberculous Mycobacterium species were detected in the one sample and two samples, respectively, among the 36 water samples by the PCR with specific primers for these bacteria. Our study indicated that DUWS in student clinical simulation laboratory can cause potential infection in students and participants. This study suggested the dental unit water line management and wearing personal protective equipment in student clinical simulation laboratory will be needed to reduce bacterial contamination.

Human Papillomavirus Prevalence and Genotype Distribution in Normal and ASCUS Specimens: Comparison of a Reverse Blot Hybridization Assay with a DNA Chip Test

  • Kim, Sunghyun;Lee, In-soo;Lee, Dongsup
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.32-39
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    • 2015
  • High-risk (HR) human papillomavirus (HPV) genotypes are strongly associated with cervical cancer, whereas other HPV genotypes are not. To identify the various HPV genotypes in clinical samples, we conducted HPV genotyping using a DNA chip test and reverse blot hybridization assay (REBA) in normal cytology samples and atypical squamous cells of undetermined significance (ASCUS) cytology samples. We also investigated the HPV infection rate and HPV genotype prevalence in women with normal cytology and ASCUS cytology. Liquid-based cytology preparations were used for the initial screening of 205 subjects with normal cytology and ASCUS cytology. The HPV infection rate was 49.8% when using the DNA chip assay and 61.0% when using the REBA test. In patients with normal cytology, the HR-HPV positive rate was 21.9% with the DNA chip assay and 43.9% with the REBA test. In contrast, 8.3% of patients with ASCUS were HR-HPV positive when using the DNA chip assay, and 13.6% were positive when tested with the REBA test. The infection rate of HR-HPV in the 40~50-year age group was significantly higher than that of the other age groups. Based on the cytological analysis of the normal and ASCUS samples, the five most prominent HPV genotypes were HPV 16, 18, 68, 33, and 58 using the DNA chip test, and they were HPV 16, 18, 53, 33, and 66 when using the REBA test. In conclusion, the findings show that the results of the REBA test are comparable to those of the DNA chip test. Most strikingly, the REBA test detected the HR-HPV genotype associated with cervical carcinoma similar to that detected with the DNA chip method. Therefore, the REBA test is a useful method to detect clinically important HR-HPV genotypes.

Development and Assessment of New RT-qPCR Assay for Detection of HIV-1 Subtypes

  • Lim, Kwanhun;Park, Min;Lee, Min Ho;Woo, Hyun Jun;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.22 no.3
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    • pp.83-97
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    • 2016
  • The measurement of viral load in HIV-1 infected patients is essential for the establishment of a therapeutic strategy. Several commercial assays have shown shortcomings in quantifying rare genotypes of HIV-1 such as minor groups of N and O. In this study, the HIV-1 RT-qPCR assay was developed. The primers and probe of HIV-1 were designed to target the pol gene and to increase the detection efficiency of various subtypes including group N and O. The HIV-1 quantitative RT-qPCR assay was assessed for its analytical performance and clinical evaluation. The LoD was determined to 33.9 IU/ml. The LoD of several subtypes including A, C, D, CRF_01AE, F, CRF_02AG, G and H, were determined to less than 40 IU/ml. The HIV-1 quantitative RT-qPCR assay was evaluated using the China National Reference Panel of HIV-1 RNA to determine the analytical performance. The results were all within the acceptable range. The clinical evaluation was performed at Hunan CDC in China. The clinical evaluation results were compared with those of the China domestic commercial kit. A significant correlation (fresh samples; $R^2=0.84$, P<0.001, frozen samples; $R^2=0.76$, P<0.001) between the two systems was observed for 64 fresh samples and 76 frozen samples with viral loads, and the Bland-Altman plot showed good agreement (98.4%, 96.1%, respectively). In conclusion, the HIV-1 quantitative RT-qPCR assay had comparable analytical performance with several commercial kits. The study provides basic data for the research of HIV-1 diagnosis and the development of P < HIV-1 molecular diagnostic assay.

Paired analysis of tumor mutation burden calculated by targeted deep sequencing panel and whole exome sequencing in non-small cell lung cancer

  • Park, Sehhoon;Lee, Chung;Ku, Bo Mi;Kim, Minjae;Park, Woong-Yang;Kim, Nayoung K.D.;Ahn, Myung-Ju
    • BMB Reports
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    • v.54 no.7
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    • pp.386-391
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    • 2021
  • Owing to rapid advancements in NGS (next generation sequencing), genomic alteration is now considered an essential predictive biomarkers that impact the treatment decision in many cases of cancer. Among the various predictive biomarkers, tumor mutation burden (TMB) was identified by NGS and was considered to be useful in predicting a clinical response in cancer cases treated by immunotherapy. In this study, we directly compared the lab-developed-test (LDT) results by target sequencing panel, K-MASTER panel v3.0 and whole-exome sequencing (WES) to evaluate the concordance of TMB. As an initial step, the reference materials (n = 3) with known TMB status were used as an exploratory test. To validate and evaluate TMB, we used one hundred samples that were acquired from surgically resected tissues of non-small cell lung cancer (NSCLC) patients. The TMB of each sample was tested by using both LDT and WES methods, which extracted the DNA from samples at the same time. In addition, we evaluated the impact of capture region, which might lead to different values of TMB; the evaluation of capture region was based on the size of NGS and target sequencing panels. In this pilot study, TMB was evaluated by LDT and WES by using duplicated reference samples; the results of TMB showed high concordance rate (R2 = 0.887). This was also reflected in clinical samples (n = 100), which showed R2 of 0.71. The difference between the coding sequence ratio (3.49%) and the ratio of mutations (4.8%) indicated that the LDT panel identified a relatively higher number of mutations. It was feasible to calculate TMB with LDT panel, which can be useful in clinical practice. Furthermore, a customized approach must be developed for calculating TMB, which differs according to cancer types and specific clinical settings.