• Annals of Laboratory Medicine
• /
• 제38권6호
• /
• pp.518-523
• /
• 2018

Background: Lipemia, a significant source of analytical errors in clinical laboratory settings, should be removed prior to measuring biochemical parameters. We investigated whether lipemia in serum/plasma samples can be removed using a method that is easier and more practicable than ultracentrifugation, the current reference method. Methods: Seven hospital laboratories in Spain participated in this study. We first compared the effectiveness of ultracentrifugation ($108,200{\times}g$) and high-speed centrifugation ($10,000{\times}g$ for 15 minutes) in removing lipemia. Second, we compared high-speed centrifugation with two liquid-liquid extraction methods-LipoClear (StatSpin, Norwood, USA), and 1,1,2-trichlorotrifluoroethane (Merck, Darmstadt, Germany). We assessed 14 biochemical parameters: serum/plasma concentrations of sodium ion, potassium ion, chloride ion, glucose, total protein, albumin, creatinine, urea, alkaline phosphatase, gamma-glutamyl transferase, alanine aminotransferase, aspartate-aminotransferase, calcium, and bilirubin. We analyzed whether the differences between lipemia removal methods exceeded the limit for clinically significant interference (LCSI). Results: When ultracentrifugation and high-speed centrifugation were compared, no parameter had a difference that exceeded the LCSI. When high-speed centrifugation was compared with the two liquid-liquid extraction methods, we found differences exceeding the LCSI in protein, calcium, and aspartate aminotransferase in the comparison with 1,1,2-trichlorotrifluoroethane, and in protein, albumin, and calcium in the comparison with LipoClear. Differences in other parameters did not exceed the LCSI. Conclusions: High-speed centrifugation ($10,000{\times}g$ for 15 minutes) can be used instead of ultracentrifugation to remove lipemia in serum/plasma samples. LipoClear and 1,1,2-trichlorotrifluoroethane are unsuitable as they interfere with the measurement of certain parameters.

• Journal of Genetic Medicine
• /
• 제12권2호
• /
• pp.96-99
• /
• 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.

• 대한임상검사과학회지
• /
• 제45권3호
• /
• pp.87-90
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제19권11호
• /
• pp.1475-1481
• /
• 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.

• Parasites, Hosts and Diseases
• /
• 제61권1호
• /
• pp.15-23
• /
• 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.

• IMMUNE NETWORK
• /
• 제2권4호
• /
• pp.202-207
• /
• 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.

• 대한임상검사과학회지
• /
• 제50권2호
• /
• pp.93-99
• /
• 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.

• 치위생과학회지
• /
• 제15권2호
• /
• pp.232-237
• /
• 2015

이 연구는 강릉원주대학교 치과대학 학생들의 임상실습을 위해 사용되고 있는 DCU에서 배출되는 물 속 종속영양세균의 수준을 평가하면서 사용빈도에 따른 세균 오염수준의 차이를 확인하고 기회 감염성 병원균의 존재를 분자생물학적 방법을 사용하여 확인하였다. 임상 실습실에서 사용되는 DCU 36개를 대상으로 초음파치석제거기에서 물 시료를 수집하여 평균 CFU/ml를 조사하고 초음파치석제거기의 한달 사용빈도에 따라 DCU를 세 집단으로 분류하여 세균오염수준을 비교하였다. 또한 수집한 물 시료에서 세균의 genomic DNA를 추출한 후 PCR 분석을 통해 기회감염성 병원균의 존재를 확인한 결과는 다음과 같다. 학생 실습에 사용한 DCU에서 수집한 물 시료의 평균 종속영양세균수준은 16,095 CFU/ml로 ADA에서 권장하는 200 CFU/ml 이하의 수준에 적합하지 않은 것을 확인하였다. 초음파 치석제거기의 한 달 사용빈도에 따라 3집단으로 나누어 CFU/ml를 조사하였을 때, 초음파치석제거기를 한 달에 1번 이상 3번 미만 사용한 DCU에서 평균 CFU/ml가 20,070 CFU/ml로 가장 높게 나타났으며, 3번 이상 사용한 유니트는 CFU/ml 평균이 8,420 CFU/ml로 가장 적게 나타났다. 3개군의 CFU/ml 차이는 통계학적으로 유의성이 있는 것을 보여주었고(p<0.05), 그 중 사용빈도가 가장 높은 군에서 유의하게 낮은 CFU/ml를 보여주었다. 치과에서 사용하는 DCU에 존재하는 기회감염성 병원균이 학생실습에 사용하는 DCU에서도 분리되었다. 36개의 genomic DNA 시료 중 1개의 시료에서 Pseudomonas species가 검출되었고, 2개의 시료에서 비결핵성 Mycobacterium species가 검출되었다. 따라서 학생실습용으로 사용되는 DCU는 학생들과 대상자에게 잠재적 감염의 원인이 될 수 있으며, 실습 전 학생들의 보호장비 착용과 실습 후 수관관리가 필요하다.

• 대한의생명과학회지
• /
• 제21권1호
• /
• pp.32-39
• /
• 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.

• 대한의생명과학회지
• /
• 제22권3호
• /
• pp.83-97
• /
• 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.