• Biomedical Science Letters
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• 제22권3호
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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.

• BMB Reports
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• 제54권7호
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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.

• Medical Lasers
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• 제9권2호
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• pp.172-178
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• 2020

Background and Objectives This study aimed at examining the effects of photobiomodulation therapy (PBMT) on glucose, cholesterol, triglycerides, and low- and high-density lipoprotein (LDL and HDL, respectively) levels in vitro. Materials and Methods A total of 38 serum samples collected in plain (n=10) and heparinized tubes (n=28) were subjected to PBMT at 60 Joules (J)/cm² for 2 min at 810 nm. The glucose and lipid profiles, cholesterol, triglycerides, LDL, and HDL of each sample was measured before and after PBMT. Results A statistically significant increase in glucose levels was observed in the PBMT-sera in 8 out of 10 samples in plain tubes. However, only two samples that were prepared in heparinized tubes showed an increase in glucose levels. The remaining heparinized samples that were exposed to PBMT presented lower glucose values. The treated sera exhibited a fluctuation in the lipid profiles after PBMT. However, high cholesterol levels were evident following PBMT. Similar trends with HDL and LDL in heparinized tubes were evident. Conclusion Together, the findings suggest that photobiomodulation exhibits an effect on glycemic and lipid profiles in vitro. Hence, the use of low-level laser therapy could have therapeutic potential. However, the differences between individual responses appear to indicate that the impact of PBMT may not always be beneficial.

• Biomedical Science Letters
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• 제24권3호
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• pp.230-238
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• 2018

Human enteric Adenovirus-41 (HuEAdV-41) causes gastroenteritis, which detected by the polymerase chain reaction (PCR) base diagnostic system for clinical, food, environmental, fish and shellfish samples. We developed improved PCR and nested PCR primer set which had high specificity, sensitivity and reduced times. In this study, we compared seventeen conditions reported in the previous study that was using the PCR based HuEAdV-41 detection system, and non-enteric Adenovirus were detected in nine conditions. The most sensitive detection condition was up to 25 copies however it took 184 minutes of PCR reaction time. In this study, the PCR primer set developed had same level of sensitivity, it reduced the time of detection for clinical, food and seafood samples to 112 minutes. Developed nested PCR primer set needed 112 minutes but detected up to approximately 1 copy. In addition, developed PCR and nested PCR primer set was validated with twenty samples of underground water at random, of which ten samples showed specific band without non-specific reaction. We expect this study will be used to diagnose HuEAdV-41 from various samples.

• Archives of Pharmacal Research
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• 제19권6호
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• pp.486-489
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• 1996

A simple, rapid, specific and sensitive method for the detection of serum hepatitis C virus (HCV) RNA using the reverse transcription-polymerase chain reaction (RT-PCR) technique without conventional RNA extraction was developed. HCV template RNA from serum was obtained by boiling the serum at $95^{\circ}C$ for 2 min, cooling rapidly in ice and removing the proteins by cetrifugation. RT-PCR amplifications including the reverse transcription and first PCR amplification were performed in one vessel containing both of reverse transcriptase and Taq DNA polymerase. The detection of HCV RNA from $10^{-3}{\mu}l$. serum was possible with this method. The suitability of this method for clinical analysis was evaluated by assaying HCV RNA in 225 patient samples including anti-HCV antibody negatives (13 samples) and positives (212 samples) by enzyme-linked immunosorbent assay test (ELISA). Detections of HCV RNA with this method were in 4 of 13 anti-HCV antibody negative samples (30.8%) and 95 of 212 positive samples (44.8%). The present method can be completed in 1 hr and has a wide range of application for the clinical utilities to determine the viral RNAS.

• Korean Journal of Clinical Laboratory Science
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• 제56권1호
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• pp.59-65
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• 2024

This study examined the fecal samples of striped field mice (Apodemus agrarius coreae) captured in Jeju Special Self-Governing Province. Fecal samples, including the colon and other intestinal organs, were collected and subjected to aerobic culture to investigate the distribution of intestinal microorganisms. Gram staining of the aerobic cultured bacterial colonies from 36 fecal samples revealed the predominant presence of gram-negative bacilli in all samples. Among the 36 samples, gram-negative bacilli were identified in 36 strains (100%), gram-positive cocci in 21 strains (58.3%), and gram-positive bacilli in 15 strains (41.7%), while no gram-negative cocci were observed. The gram-negative bacilli cultured from the 36 samples were identified using the Vitek 2 system, and all were determined to be Escherichia coli (E. coli) strains. In addition, one sample was concurrently identified with E. coli and Enterobacter cloacae strains. The antimicrobial susceptibility testing for the identified E. coli strains did not include all antibiotics, but one strain exhibited intermediate resistance to cefoxitin. No pathogenic bacteria were present in the fecal samples of the scrub typhus-infected rodents, which are vectors for chigger-borne diseases affecting humans and animals.

• Biomedical Science Letters
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• 제22권3호
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• pp.98-106
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• 2016

Investigation of human papillomavirus (HPV) in archival formalin-fixed paraffin-embedded (FFPE) material is important for understanding cervical carcinogenesis. The objective of the present study was to identify the high risk HPVs (HR-HPVs) using HPV E6/E7 mRNA testing from archival tissues in cervical cancer and the relation to HR-HPVs genotypes in paired cervical exfoliated cells. HPV E6/E7 mRNA testing and DNA chip testing were performed in 79 paired cervical FFPE tissues and exfoliated cells from women with histologically confirmed squamous cell carcinoma and adenocarcinoma. Overall agreement in HR-HPVs detection from FFPE samples and cytology samples were 98.5% in HPV 16, 100% in HPV 18, HPV 31, HPV 33, HPV 58, HPV 66, and HPV 68. Type-specific agreement between FFPE samples and cytology samples was 89.1% in HPV positive, 93.5% in HPV 16 and more than 70% in the other HR-HPVs. In conclusion, HR-HPVs were reliably detected in paired FFPE and cytology samples with some variation in type-specific detection.

• Asian Pacific Journal of Cancer Prevention
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• 제17권2호
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• pp.673-676
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• 2016

Background: In the maxillofacial region, giant cell granulomas occur in 2 clinical forms, central and peripheral. Despite histopathological similarity between these 2 forms totally different clinical behaviors have been reported. The present study was undertaken to compare mast cell and vascular concentrations in these pathologic lesions. Materials and Methods: In this cross-sectional descriptive study, 20 pathological samples of central giant cell granuloma (CGCG) and 20 samples of peripheral giant cell granuloma (PGCG) were selected and examined through toluidine blue staining for mast cell assessment and immunohistochemical staining by VEGEF antibody for comparing the number of mast cells. T-test, chi-squared test and backward multivariate linear regression were used for statistical analysis using SPSS 20. Statistical significance was set at P<0.05. Results: This study showed significantly greater VEGF expression and mast cell concentrations in CGCG compared to PGCG cases. Also there was a significant correlation between VEGF expression and the concentration of mast cells. No relation was found between age, sex and site of the lesion and concentration of mast cells or VEGF expression. Conclusions: It is feasible that higher concentrations of mast cells in CGCG versus PGCG samples might lead to more aggressive clinical behavior via vascular proliferation and angiogenesis. However, other biologic mechanisms should be considered in this situation.

• The Journal of the Korean Society for Microbiology
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• 제21권1호
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• pp.107-111
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• 1986

An outbreak of Pontiac fever was reported in Seoul in 1984, but legionnaires disease was not known in Korea yet. Our knowledge on the presence or abscence of Legionella in cooling tower. which is the main source of the infection. is very limited. In this study an attempt was made to determine the presence of Legionella in cooling towers during June-September. and in the sputum specimens for routine bacteria culture, which was taken during July-August 1985. Among the 83 water samples 6 yielded L. pneumophila serogroup 1, while none of the 189 sputum samples yielded growth of Legionella. It is concluded that legionellosis can occur in Korea and if it happens it is most likely due to L. pneumophila serogroup 1.

• The Journal of the Korean Society for Microbiology
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• 제19권1호
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• pp.73-77
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• 1984

Vibrio vulnificus is an organism capable of causing septicemia and wound infection in compromised patients. The source of infection is known to be raw oysters and others. The prevalence of the organism in Korean sea water and shellfish is not known. The authors surveyed shellfish and other specimens obtained mainly from a market in Seoul and from ones in Inchon. Five cultures of V. vulnificus were isolated from oyster and clam samples. Two isolates had typical characteristics of the strains isolated from patients, i.e., definite hemolysis and typical biochemical reactions. However, other 2 isolates were sucrose positive, although the identity were confirmed by Center for Disease Control. We do not know wether such strains are pathogenic or not. For the isolation of V. vulnificus from environmental samples, TCBS agar and VV agar were not very selective or differential. We isolated our strains with the use of OF-lactose agar and SPS agar. However OF-lactose agar did not support good growth of V. vulnificus, while SPS agar did not suppress other vibrios.