Herein, we report a case in which cholestasis caused discrepancy in high-density lipoprotein (HDL)-cholesterol levels measured using various methods. The discrepancy in HDL-cholesterol level originated from the abnormal increase in the level of an unusual lipoprotein, apo E-rich HDL, in the patient's serum. An abnormal slow alpha-migrating lipoprotein was observed on agarose gel electrophoresis, and an abnormal large-sized HDL was observed in a lipoprotein subfraction study. The level of apolipoprotein E was elevated.
Apolipoproteins ; Cholestasis ; Electrophoresis, Agar Gel ; Lipoproteins

Herein, we report a case in which cholestasis caused discrepancy in high-density lipoprotein (HDL)-cholesterol levels measured using various methods. The discrepancy in HDL-cholesterol level originated from the abnormal increase in the level of an unusual lipoprotein, apo E-rich HDL, in the patient's serum. An abnormal slow alpha-migrating lipoprotein was observed on agarose gel electrophoresis, and an abnormal large-sized HDL was observed in a lipoprotein subfraction study. The level of apolipoprotein E was elevated.
Apolipoproteins ; Cholestasis ; Electrophoresis, Agar Gel ; Lipoproteins

BACKGROUND: Recently biphasic agarose gel electrophoresis method using Chol/Trig ComboTM for simultaneous detection of cholesterol and triglyceride on lipoprotein fractions has been developed to facilitate the classification and interpretation of abnormal lipoprotein patterns of patients with hyperlipidemia. The author analysed serum lipid profiles using Chol/Trig ComboTM in the patients of OPD in Department of Internal Medicine. METHODS: Measurement of serum cholesterol and triglyceride using enzymatic method was performed in the sera of 415 patients from April, 2006 to July, 2006. Simultaneously, we electrophoresed serum cholesterol and triglyceride using Chol/Trig ComboTM with analysis software (ED BANK, Helena Laboratories, Saitama, Japan). RESULTS: According to ATP III guideline, we set up standard cholesterol as 200 mg/dL and triglyceride as 150 mg/dL, respectively, and the patients were classified into control, hypercholesterolemic, hypertriglyceridemic and hypercholesterolemic/hypertriglyceridemic groups, respectively. Hypercholesterolemic group had higher HDL and LDL, hypertriglyceridemic group had higher HDL, VLDL and the rates of positive modified LDL, hypercholesterolemic/hypertriglyceridemic group have higher VLDL and the rates of positive modified LDL than the control. Otherwise, the rates of positive modified LDL were 19.8% in the control, 23.6% in the hypercholesterolemic group, 50.9% in hypertriglyceridemic group, and 39.3% in hypercholesterolemic/hypertriglyceridemic group. CONCLUSIONS: The frequency of the rates of positive modified LDL and VLDL was higher in the hypertriglyceridemic group than those in the control.
Adenosine Triphosphate ; Cholesterol ; Electrophoresis ; Electrophoresis, Agar Gel ; Humans ; Hyperlipidemias ; Lipoproteins

BACKGROUND: Recently biphasic agarose gel electrophoresis method using Chol/Trig ComboTM for simultaneous detection of cholesterol and triglyceride on lipoprotein fractions has been developed to facilitate the classification and interpretation of abnormal lipoprotein patterns of patients with hyperlipidemia. The author analysed serum lipid profiles using Chol/Trig ComboTM in the patients of OPD in Department of Internal Medicine. METHODS: Measurement of serum cholesterol and triglyceride using enzymatic method was performed in the sera of 415 patients from April, 2006 to July, 2006. Simultaneously, we electrophoresed serum cholesterol and triglyceride using Chol/Trig ComboTM with analysis software (ED BANK, Helena Laboratories, Saitama, Japan). RESULTS: According to ATP III guideline, we set up standard cholesterol as 200 mg/dL and triglyceride as 150 mg/dL, respectively, and the patients were classified into control, hypercholesterolemic, hypertriglyceridemic and hypercholesterolemic/hypertriglyceridemic groups, respectively. Hypercholesterolemic group had higher HDL and LDL, hypertriglyceridemic group had higher HDL, VLDL and the rates of positive modified LDL, hypercholesterolemic/hypertriglyceridemic group have higher VLDL and the rates of positive modified LDL than the control. Otherwise, the rates of positive modified LDL were 19.8% in the control, 23.6% in the hypercholesterolemic group, 50.9% in hypertriglyceridemic group, and 39.3% in hypercholesterolemic/hypertriglyceridemic group. CONCLUSIONS: The frequency of the rates of positive modified LDL and VLDL was higher in the hypertriglyceridemic group than those in the control.
Adenosine Triphosphate ; Cholesterol ; Electrophoresis ; Electrophoresis, Agar Gel ; Humans ; Hyperlipidemias ; Lipoproteins

BACKGROUND: Polymerase chain reacation (PCR)-based methods have been described for rapid detection and identification of Candida spp. Multiplex PCR assay was developed using internal transcribed spacers and topoisomerase II gene for the accurate identification of Candida species. METHODS: We designed Dual Specificity Oligo (DSO) primers for multiplex PCR. Multiplex PCR was followed by agarose gel electrophoresis to test 8 type strains (C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae, C. dubliniensis) and 96 clinical isolates (C. albicans 51 isolates, C. parapsilosis 10 isolates, C. glabrata 10 isolates, C. tropicalis 9 isolates, C. krusei 6 isolates, C. guilliermondii 5 isolates, C. lusitaniae 5 isolates) of Candida spp. RESULTS: With multiplex PCR using DSO primers, the eight Candida type strains each could be easily differentiated and all 96 clinical isolates were identified as the same species as were identified by the conventional method. CONCLUSION: Multiplex PCR followed by electrophoresis can be useful for the simple and rapid identification of Candida species in routine laboratories.
Candida* ; DNA Topoisomerases, Type II ; Electrophoresis ; Electrophoresis, Agar Gel ; Multiplex Polymerase Chain Reaction* ; Sensitivity and Specificity

OBJECTIVETo analyse clinical feasibility of two electrophoresis procedures of seminal plasma proteins, agarose gel electrophoresis and SDS-agarose gel electrophoresis.

METHODSSixty-nine semen samples were examined and classified into three groups: the asthenozoospermia (n = 22), the asthenoteratozoospermia (n = 19), and the relative normal group (n = 28) with normal routine and special test results, according to WHO routine and special test criterion. Then, the seminal plasma protenis were separated by two different electrophoresis, with SDS-agarose and agarose support medium, the buffer pH 7.0 and 9.2 respectively. The agarose gel electrophoresis was done under various sample loading time, motion power and staining modules. The completed gels were scanned and compared the each other statistically.

RESULTSSeminal plasma proteins can be separated into 4 strips by SDS-agarose gel electrophoresis with acid crystal violet, and the strips were diffusion and with dark background. However, 6 clear strips named A, B, C, D, E, and F can be obtained by agarose gel electrophoresis with 6 min. After samples were loaded and stained by amidoblack, there showed appropriate spaces among strips, and it was very easy to scan the drying gel by a densitometer. Using agarose gel electrophoresis, the statistical difference in strip C and E was shown between the asthenozoospermia and the relative normal group, and between the asthenozoospermia and the asthenoteratozoospermia, however, not between the relative normal and the asthenoteratozoospermia group. Moreover, the samples in the relative normal group with normal routine and special test results were in different electrophoresis maps.

CONCLUSIONAgarose gel electrophoresis of seminal plasma proteins with buffer pH 9.2, 6 min. sample loading and amidoblack stain was a simple, fast and fit technique for clinic.

Adult ; Electrophoresis, Agar Gel ; methods ; Electrophoresis, Polyacrylamide Gel ; methods ; Humans ; Male ; Proteins ; analysis ; Semen ; chemistry ; Staining and Labeling

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) has a heteroresistant nature, so methicillin resistance is influenced by various culture conditions, such as temperature, incubation time, and NaCl content in the medium. Mueller Hinton (MH) agar containing 2% NaCl and mannitol salt agar (MSA) with oxacillin disk were evaluated for the detection of methicillin resistance. METHODS: Disk diffusion test on plain Mueller- Hinton (MH) agar, 2% NaCl MH agar, and MSA with 1 microgram oxacillin disk was performed in 70 Stap hylococcus aureus isolates. Oxacillin MIC was determined by E-test. As a gold standard of methicillin resistance, mecA gene was amplified by PCR and detected by agarose gel electrophoresis. RESULTS: Plain MH agar could not detect heterogeneous resistance in 12 S. aureus isolates (18%), but 2% NaCl MH agar and MSA could correctly detect homogeneous and heterogeneous resistance. S. aureus isolates from stool have as much as 48% heterogeneous resistance, while those from non-stool specimen have 5%. CONCLUSION: 2% NaCl and MSA can be used reliably for accurate susceptibility testing of methicillin resistance in routine laboratory.
Agar* ; Diffusion ; Electrophoresis, Agar Gel ; Mannitol* ; Methicillin Resistance* ; Methicillin* ; Methicillin-Resistant Staphylococcus aureus ; Oxacillin ; Polymerase Chain Reaction ; Staphylococcus aureus* ; Staphylococcus*

BACKGROUND: The determination of apo E polymorphism through the phenotyping is not suitable for large studies in the clinical laboratories because of various problems. So apo E genotyping has been developed. We present a method of apo E genotyping using a modified amplification refractory mutation system(ARMS) technique. METHODS: We used four primers to detect the region containing two mutation points coding amino acid residues 112 and 158 and have developed a method of apo E genotyping by the modified ARMS technique. Apo E genotyping were performed by both the modified ARMS technique and the INNO-LiPA Apo E kit(Innogenetics, Belgium) with the following frequency distribution: Epsilon2/2(n=10), Epsilon3/3(n=15), Epsilon4/4(n=7), Epsilon2/3(n=9), Epsilon2/4(n=12), Epsilon3/4(n=13). RESULTS: All the samples gave the correct and clear amplification patterns. Modified ARMS correctly distinguished among the six apo E genotypes. The apo E genotypes determined by both methods for every specimen studied were in complete agreement. CONCLUSIONS: This modified ARMS technique involved only two stages: PCR and agarose gel electrophoresis. Since apo E genotyping by the modified ARMS is reliable, simple to perform, less time consuming, and not expensive, we conclude that it is suitable for large sample studies in the clinical laboratories.
Apolipoproteins E ; Apolipoproteins* ; Arm* ; Clinical Coding ; Electrophoresis, Agar Gel ; Genotype ; Polymerase Chain Reaction*

PURPOSE: The success of basic molecular research using biospecimens strongly depends on the quality of the specimen. In this study, we evaluated the effects of delayed freezing time on the stability of DNA and RNA in fresh frozen tissue from patients with colorectal cancer. METHODS: Tissues were frozen at 10, 30, 60, and 90 minutes after extirpation of colorectal cancer in 20 cases. Absorbance ratio of 260 to 280 nm (A(260)/A(280)) and agarose gel electrophoresis were evaluated. In addition, the RNA integrity number (RIN) was assayed for the analysis of the RNA integrity. RESULTS: Regardless of delayed freezing time, all DNA and RNA samples revealed A(260)/A(280) ratios of more than 1.9, and all DNA samples showed a discrete, high-molecular-weight band on agarose gel electrophoresis. The RINs were 7.53 +/- 2.04, 6.70 +/- 1.88, 6.47 +/- 2.58, and 4.22 +/- 2.34 at 10, 30, 60, and 90 minutes, respectively. Though the concentration of RNA was not affected by delayed freezing, the RNA integrity was decreased with increasing delayed freezing time. CONCLUSION: According to the RIN results, we recommend that the collection of colorectal cancer tissue should be done within 10 minutes for studies requiring RNA of high quality and within 30 minutes for usual RNA studies.
Colorectal Neoplasms ; DNA ; Electrophoresis, Agar Gel ; Freezing ; Humans ; Quality Control ; RNA ; Tissue Banks

BACKGROUND: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(Polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. METHODS: We used the InstaGene(TM) DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1, 245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2, 536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. RESULTS: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value.. CONCLUSION: Even though both methods had lower possibility of cross contamination, shorter time requrirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usfulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.
DNA* ; Electrophoresis, Agar Gel ; Endopeptidase K ; Ion Exchange* ; Microwaves ; Polymerase Chain Reaction* ; Sensitivity and Specificity ; Sputum ; Tuberculosis*