• Parasites, Hosts and Diseases
• /
• 제39권2호
• /
• pp.171-176
• /
• 2001

Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), meroxoite surface protein (MSP-1), apical merozoite antigen (AMA- 1), serine repeat antigen (SERA), and exported antigen (EXP- 1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using S antigens (91.9%) rather than using each antigen solely (55.6 - 80%), a combination of 2 (76.3 - 87.5%), 3 (85.6 - 90.6%), or 4 antigens (89.4 - 91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.

• 농촌의학ㆍ지역보건
• /
• 제16권1호
• /
• pp.70-78
• /
• 1991

Antibody changes in experimental anisakiasis were observed by ELlSA and SDS-PAGE/EITB using various antigens : whole worm extract antigen(WWE), somatic antigen(SOM), excretory-secretory antign(ES), and hemoglobin antigen(HB) of Anisakis Type 1. The results obtained were as follows. l) Serum levels of IgG antibody by ELISA increased from 1st week of infection and reached their maximum titer at 5th week after infection, and decreased gradually thereafter. 2) The best result expressed as positive/negative ratio could be obtained when ES antigen was used. 3) Silver stained SDS-PAGE of each antigen showed at least 20 protein bands : In WWE, 286, 278, 262, 38, 18 Kd bands ; In SOM, 38 Kd band : In ES, 286, 65, 13 Kd bands ; In Hb, 61, 55, 38, 28, 26, 22, 20, 16, 15 Kd bands iepntibied as were major bands. 4) By EITB using WWE, Serum antibody recognized major protein with molecular weight of 86 Kd and 16 Kd. Using ES, 69, 59, 16 Kd bands were observed and using Hb, 28 Kd band was observed as specific band. In conclusion, excretory-secretory antigen(ES) of Anisakis larvae was most usable for ELISA.

• 대한수의학회지
• /
• 제41권1호
• /
• pp.43-49
• /
• 2001

As compared with reaction of antibody for sonicated antigen of Brucella abortus strain RB51 and 1119-3 by Western blot analysis, Brucella field positive sera was detected strong reaction at 40~80 kDa LPS of strain 1119-3, but detected very weak reaction at strain RB51 partly. Otherwise, as we analyzed major immunogen of RB51 by antisera bled periodically during 6 months after RB51 vaccination. we detected strong immunological reaction at 17, 18 and 8 kDa antigen of RB51. Especially, reaction of 8 kDa antigen by Western blot coincided with reaction of dot-blot assay in RB51-antibody detection method. We also compared with reaction of field sera by STAT(standard tube agglutination test), dot-blot assay and Western blot (reaction of 8 kDa antigen of strain RB51). 16 sera of 4~5 months after RB51 vaccination are all negative by STAT, and 12 field brucellosis positive serum are all positive, and also 12 of 16 sera vaccinated RB51 are positive by dot-blot assay and reaction of 8kDa antigen by Western blot. but 1 of 15 Brucellosis negative sera reacted nonspecifically dot-blot assay.

• 한국동물위생학회지
• /
• 제40권1호
• /
• pp.61-66
• /
• 2017

Calf diarrhea is a disease experienced by almost all of calves after birth and is one of the representative causes of damage to farmers due to mass mortality and of economic losses to them by inhibiting normal growth. In this study, we conducted quick detection of etiologic agents of diarrhea by using a rapid diagnostic kit to multiply diagnose antigens of five etiologic agents of calf diarrhea (rotavirus, coronavirus, Escherichia coli, Cryptosporidium, Giardia) in Hanwoo (Korean native cattle) calves. When the positive antigen proportion of the calf diarrheal feces for each farm was analyzed, rotavirus, coronavirus, Escherichia coli, Cryptosporidium, and Giardia showed antigen positive rates of 0~67%, 0~20%, 0~60%, 0~20%, and 0~67%, respectively. With regard to the antigen positive rate by age in days after birth, 1-week-old calves showed the antigen positive rate of 20% in rotavirus and 20% in Giardia, and 2-week-old calves showed that of 50% in rotavirus. In addition, 4-week-old calves showed the antigen positive rate of 10% in rotavirus, 10% in coronavirus, 10% in Escherichia coli, and 30% in Giardia, and 8-week-old calves showed the antigen positive rate of 17% in coronavirus, 50% in Escherichia coli, 17% in Cryptosporidium, and 33% in Giardia. Based on the results of this study, the etiologic agents of diarrhea in Hanwoo calves for each farm are widely distributed. Although younger than 2-week-old calves were strongly positive for rotavirus, older than 4-week-old calves were highly positive for Giardia and Escherichia coli. In conclusion, we considered that a rapid diagnostic kit is an effective method for quick detection of etiologic agents and would be helpful for cattle farmers and veterinarians to select appropriate therapeutic method.

• Perspectives in Nursing Science
• /
• 제7권1호
• /
• pp.50-54
• /
• 2010

The studies of M. Colombo (1989) and W. Lange (1992) showed that 30~40% of people became chronic after suffering from hepatitis B virus (HBV) and C virus (HCV) infection, and about 50% of the chronic cases transformed into primary liver cancer. There have been few studies done in Mongolia on hepatitis infection among health professionals, particularly in nurses. In a study done by Chimedsuren (8), the study showed that 19.4% of people with identified surface hepatitis B antigen (HBsAg) and antibodies to hepatitis C virus and 8% of people with the identified nucleotide of RNA for the hepatitis C virus (polymerase chain reaction) had an acute form of hepatitis C. Studies on the hepatitis virus genome damaging effect on liver cells showed that genotype 8 (A, B, C, D, E, F, G, TTV) had the most damaging effect on liver cells (Hahn and Faeka, 2007). Several studies have shown a relationship between hepatitis B virus infection and a lack of compliance regarding safety regulations and rules by medical personnel. Results of a study from the Maternal and Child Health Research Center showed that tests done to detect hepatitis B virus antigen and antibodies to C virus did not reveal anything. Both antigen and antibodies in 69% cases did not show, and separately, B virus and antibodies to hepatitis C virus were identified in 13% and 9%, respectively. Results of the tests taken from health personnel in Shastin Central Hospital showed that in 76% of the cases, the B virus antigen with C virus antibodies was not identified. In 8% of the cases, the B virus antigen was present on its own. The combination of B the virus antigen and C virus antibodies were present in 8% of nurses and doctors, respectively. 82% of the cases had negative results for the detection of a combination of B virus antigen and C virus antibodies taken from health personnel from the State Central Clinical Hospital whereas the B virus antigen and C virus antibodies by themselves were present in 7% and 14% of the cases, respectively. Combined cases of the B virus antigen and C virus antibodies were identified in 4% of the personnel. Results of the tests taken from the health personnel in the Hospital of the Ministry of Justice and Internal Affairs showed that in 79% of the cases, the B virus antigen with C virus antibodies were not identified. Separately, the B virus and antibodies to hepatitis C virus were identified in 8% and 13% of the cases, respectively.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권9호
• /
• pp.3867-3870
• /
• 2015

Background: To explore whether combined detection of serum tumor markers (CEA, CA72-4, CA19-9 and TSGF) improve the sensitivity and accuracy in the diagnosis of gastric cancer (GC). Materials and Methods: An automatic chemiluminescence immune analyzer with matched kits were used to determine the levels of serum CEA, CA72-4, CA19-9 and TSGF in 45 patients with gastric cancer (GC group), 40 patients with gastric benign diseases (GBD group) hospitalized in the same period and 30 healthy people undergoing a physical examination. The values of those 4 tumor markers in the diagnosis of gastric cancer was analyzed. Results: The levels of serum CEA, CA72-4, CA19-9 and TSGF of the GC group were higher than those of the GBD group and healthy examined people and the differences were significant (P<0.001). The area under receiver operating characteristic (ROC) curves for single detection of CEA, CA72-4, CA19-9 and TSGF in the diagnosis of GC was 0.833, 0.805, 0.810 and 0.839, respectively. The optimal cutoff values for these 4 indices were 2.36 ng/mL, 3.06 U/mL, 5.72 U/mL and 60.7 U/mL, respectively. With combined detection of tumor markers, the diagnostic power of those 4 indices was best, with an area under the ROC curve of 0.913 (95%CI 0.866~0.985), a sensitivity of 88.9% and a diagnostic accuracy of 90.4%. Conclusions: Combined detection of serum CEA, CA72-4, CA19-9 and TSGF increases the sensitivity and accuracy in diagnosis of GC, so it can be regarded as the important means for early diagnosis.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권7호
• /
• pp.3087-3091
• /
• 2014

Purpose: To determine the utility of digital rectal examination (DRE), serum total prostate specific antigen (tPSA) estimation, and transrectal ultrasound (TRUS) for the detection of prostate cancer (PCa) in men with lower urinary tract symptoms (LUTS). Materials and Methods: All patients with abnormal DRE, TRUS, or serum tPSA >4ng/ml, in any combination, underwent TRUS-guided needle biopsy. Eight cores of prostatic tissue were obtained from different areas of the peripheral prostate and examined histopathologically for the nature of the pathology. Results: PCa was detected in 151 (50.3%) patients, remaining 149 (49.7%) showed benign changes with or without active prostatitis. PCa was detected in 13 (56.5%), 9 (19.1%), 26 (28.3%), and 103 (74.6%) of patients with tPSA <4 ng/ml, 4-10 ng/ml, 10-20 ng/ml and >20 ng/ml respectively. Only 13 patients with PCa had abnormal DRE and TRUS with serum PSA <4 ng/ml. The detection rate was highest in patients with tPSA >20 ng/ml. The association between tPSA level and cancer detection was statistically significant (p<0.01). Among 209 patients with abnormal DRE and raised serum PSA, PCa was detected in 128 (61.2%). Conclusions: The incidence of PCa increases with increasing serum level of tPSA. The overall screening and detection rate can be further improved by using DRE, TRUS and TRUS-guided prostate needle biopsies.

• 약학회지
• /
• 제49권3호
• /
• pp.217-224
• /
• 2005

Human Immunodeficiency Virus type 1 (HIV-l) based lentivirus vector has demonstrated great potential as gene therapy vectors mediating efficient gene delivery and long-term transgene expression in both dividing and nondividing cells. However, for clinical studies it must be confirmed that vector preparations are safe and not contaminated by replication competent lentivirus (RCL) related to the parental pathogenic virus, HIV-l. In this study, we would like to establish the method for titration and RCL detection of lentivirus vector. The titration was determined by vector expression containing the green fluorescent protein, GFP in transduced cells. The titer was $1{\times}10^7$ Transducing Unit/ml in the GFP expression assay and $8.9{\times}10^7$ molecules/ml in the real-time PCR. Also, for the detection of RCL, we have used a combination method of PCR and p24 antigen detection. First, PBS/psi and VSV-G region in the genomic DNA of transduced cells was detected by PCR assay. Second, transfer and expression of the HIV-1 gag gene was detected by p24 ELISA. In an attempt to amplify any RCL, the transduced cells were cultured for 3 weeks (amplification phase) and the supernatant of amplified transduced cell was used for the second transduction to determine whether a true RCL was present (indicator phase). Analysis of cells and supernatant at day 6 in indicator phase were negative for PBS/psi, VSV-G, and p24 antigen. These results suggest that they are not mobilized and therefore there are no RCL in amplification phase. Thus, real-time PCR is a reliable and sensitive method for titration and RCL detection of lentivirus vector.

• Journal of applied mathematics & informatics
• /
• 제5권3호
• /
• pp.785-796
• /
• 1998

In this paper we propose a model of HIv population through method of phases with non-Markovian evolution of immi-gration. The analysis leads to an explicit differnetial equations for the generating functions of the total population size. The detection process of antibodies (against the antigen of virus) is analysed and an explicit expression for the correlation functions are provided. A measure of bunching is also introduced for some particular choice of parameters.

• Journal of Microbiology and Biotechnology
• /
• 제17권12호
• /
• pp.1955-1964
• /
• 2007

A recent report showed that analysis of CD154 expression in the presence of the secretion inhibitor Brefeldin A (Bref A) could be used to assess the entire repertoire of antigen-specific $CD4^+\;T$ helper cells. However, the capacity of intracellular CD154 expression to identify antigen-specific $CD8^+\;T$ cells has yet to be investigated. In this study, we compared the ability of intracellular CD154 expression to assess antigen-specific $CD8^+\;T$ cells with that of accepted standard assays, namely intracellular cytokine IFN-${\gamma}$ staining (ICS) and MHC class I tetramer staining. The detection of intracellular CD154 molecules in the presence of Bref A reflected the kinetic trend of antigen-specific $CD8^+\;T$ cell number, but unfortunately showed less sensitivity than ICS and tetramer staining. However, ICS levels peaked and saturated 8 h after antigenic stimulation in the presence of Bref A and then declined, whereas intracellular CD154 expression peaked by 8 h and maintained the saturated level up to 24 h post-stimulation. Moreover, intracellular CD154 expression in antigen-specific $CD8^+\;T$ cells developed in the absence of $CD4^+\;T$ cells changed little, whereas the number of IFN-${\gamma}$-producing $CD8^+\;T$ cells decreased abruptly. These results suggest that intracellular CD154 could aid the assessment of antigen-specific $CD8^+\;T$ cells, but does not have as much ability to identify heterogeneous $CD4^+\;T$ helper cells. Therefore, the combined analytical techniques of ICS and tetramer staining together with intracellular CD154 assays may be able to provide useful information on the accurate phenotype and functionality of antigen-specific $CD8^+\;T$ cells.