The followings are the results for external quality assessment (EQA) in immunoserology for 2007: 1. Evaluation of EQA was done in 2 trials in May and December, about 99% of laboratories participating average 7.8 items. The results were collected via internet for the first time and 96~98% of laboratories have sent their results via internet. 2. All the specimens for Immunoserology in EQA were delivered refrigerated, being received within 48 hours after sending. 3. Commercial controls, MASR Immunology Control from Medical Analysis Systems (Camarillo, CA, USA) were used to assure the quality of quantitative results of C-reactive protein (CRP), rheumatoid factor (RF) and anti- streptolysin O (ASO) tests, and the RF results of MASR Immunology Control were variable depending on the reagents used. 4. The laboratories using immunochromatography assay (ICA) were increased, however, many laboratories using ICA reported falsely negative for the positive specimens. The sensitivity of ICA test kits as well as various factors influencing the ICA results should be evaluated. 5. The HBsAg results of the ACCURUN 1R Multi-Marker Positive Control (Boston Biomedica Inc. USA) were falsely reported as negative in some laboratories using arbitrarily determined cutoff. 6. Standardization of methods including calibrators for quantitative results should be required for the harmonization of results.
Bacterial Proteins ; C-Reactive Protein ; Hepatitis B Surface Antigens ; Immunochromatography ; Indicators and Reagents ; Internet ; Korea ; Nephelometry and Turbidimetry ; Rheumatoid Factor ; Streptolysins

The followings are the results for external quality assessment (EQA) in immunoserology for 2007: 1. Evaluation of EQA was done in 2 trials in May and December, about 99% of laboratories participating average 7.8 items. The results were collected via internet for the first time and 96~98% of laboratories have sent their results via internet. 2. All the specimens for Immunoserology in EQA were delivered refrigerated, being received within 48 hours after sending. 3. Commercial controls, MASR Immunology Control from Medical Analysis Systems (Camarillo, CA, USA) were used to assure the quality of quantitative results of C-reactive protein (CRP), rheumatoid factor (RF) and anti- streptolysin O (ASO) tests, and the RF results of MASR Immunology Control were variable depending on the reagents used. 4. The laboratories using immunochromatography assay (ICA) were increased, however, many laboratories using ICA reported falsely negative for the positive specimens. The sensitivity of ICA test kits as well as various factors influencing the ICA results should be evaluated. 5. The HBsAg results of the ACCURUN 1R Multi-Marker Positive Control (Boston Biomedica Inc. USA) were falsely reported as negative in some laboratories using arbitrarily determined cutoff. 6. Standardization of methods including calibrators for quantitative results should be required for the harmonization of results.
Bacterial Proteins ; C-Reactive Protein ; Hepatitis B Surface Antigens ; Immunochromatography ; Indicators and Reagents ; Internet ; Korea ; Nephelometry and Turbidimetry ; Rheumatoid Factor ; Streptolysins

OBJECTIVETo prepare pneumolysin as a new protein carrier of vaccine against otitis media with genetic engineering technology and establish the base of the study on pneumococcal conjugative vaccines.

METHODSGenomic DNA was isolated from streptococcus pneumoniae. A pair of primers which included two restriction sites was designed based on the published pneumolysin gene sequence. The pneumolysin gene was amplified from pneumococcal DNA with PCR technology. The restriction enzyme digested fragment was linked into the cloning vector PET-28a and the recombinant plasmid DNA containing pneumolysin was then transfected into host cell E. coli JM109 (DE3).

RESULTSDNA fragments were subcloned to construct the complete pneumolysin gene by a conventional coning and PCR. The inserted pneumolysin gene sequence was confirmed by DNA sequencing and the pneumolysin protein was successfully expressed. The relative molecular mass of the expressed product was 52 000. The expressed product amounted to 8% of the total host cell protein.

CONCLUSIONSThe pneumolysin gene was successfully cloned into host cell using genetic engineering technology. The recombinant pneumolysin was expressed and purified for preparation. This work laid a foundation of the preparation of pneumococcal conjugative vaccines.

Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Genetic Engineering ; Genetic Vectors ; Plasmids ; Pneumococcal Vaccines ; genetics ; Streptococcus pneumoniae ; genetics ; Streptolysins ; biosynthesis ; genetics

BACKGROUND: Antistreptolysin O (ASO) is very useful as an indicator of recent streptococcal infections and their sequelae, such as rheumatic fever and acute glomerulonephritis. To interpret single ASO level of patients, the upper limit of normal (ULN) ASO from the same age group in the area should be known. As Streptococcus pyogenes infections are quite common in elementary school, we measured ASO and analyzed them by the results of throat culture to determine upper limit of normal ASO of school children in Chinju area. METHODS: ASO concentrations were measured quantitatively by nephelometry on 436 sera of healthy elementary school children in Chinju area. Throat cultures were taken at the same time to evaluate the relationship between ASO concentrations and throat culture results, including serogroup, colony forming units (CFU), and M types. RESULTS: The mean ASO concentration was 285IU/ml and the upper limit of normal ASO was 433IU/ml. The ASO levels were even (253-285IU/ml) through whole school grades except the 5th grade (350IU/ml). Not only the carriers of group A streptococci, but also those of group C or group G streptococci had higher ASO levels. The children from whom more than 10 CFU of S. pyogenes were isolated showed higher ASO levels than those who had less than 10 CFU. The ASO levels were higher in M type 6 or 22 compared to M type 12 or 28. CONCLUSIONS: The upper limit of normal ASO of children in Chinju was 433IU/ml, that is between Seoul(326IU/ml) and Chungnam (499IU/ml). The children who had more than 10 CFU tended to have higher ASO levels, which indicate asymptomatic infections, are associated with burden of bacteria. Group C or group G streptococci may induce serum response like group A streptococci. Certain M types may be implicated as strong producer of streptolysin O.
Antistreptolysin* ; Asymptomatic Infections ; Bacteria ; Child ; Chungcheongnam-do ; Glomerulonephritis ; Gyeongsangnam-do ; Humans ; Nephelometry and Turbidimetry ; Pharynx ; Rheumatic Fever ; Stem Cells ; Streptococcal Infections ; Streptococcus pyogenes ; Streptolysins

BACKGROUND: Toll-like receptors (TLRs) are expressed by human epidermal keratinocytes and are involved in immune responses. OBJECTIVE: The goal of this was to investigate the expression of TLR2 in response to bacterial antigens, cytokines, and different calcium concentrations. METHODS: The expression of TLR2 was assessed after stimulation by lipoteichoic acid (LTA) and streptolysin O (SLO). In addition, TLR2 expression was evaluated after treatment with IFN-gamma and TNF-alpha, and different concentrations of calcium. The expression levels of TLR2 mRNA and protein were studied using RT-PCR and Western blot analysis. RESULTS: Cultured human epidermal keratinocytes constitutively expressed TLR2 and the expression was stimulated by LTA and SLO; in addition, IFN-gamma and TNF-alpha upregulated TLR2 expression. However, the changes in TLR2 expression associated with the calcium concentrations were insignificant. CONCLUSION: TLR2 expression increased with the concentration and duration of bacterial pathogens and this increase was amplified by several cytokines, from activated keratinocytes and other cells.
Antigens, Bacterial ; Bacterial Proteins ; Blotting, Western ; Calcium ; Cytokines ; Humans ; Keratinocytes ; Lipopolysaccharides ; RNA, Messenger ; Streptolysins ; Teichoic Acids ; Toll-Like Receptor 2 ; Toll-Like Receptors ; Tumor Necrosis Factor-alpha

This study is designed to evaluate the immune status of schoolchildren with respect to Streptococcus pyogenes, and to ascertain the usefulness of antideoxyribonuclease B (ADNase B). Antistreptolysin O (ASO) and ADNase B concentrations were measured quantitatively in 266 serum samples from healthy elementary school children in Seoul. Simultaneously, throat cultures were taken in order to isolate S. pyogenes and other beta-hemolytic streptococci (BHS). The upper limits of the normal (ULN) concentration of ASO and ADNase B were 326 IU/mL, and 362 IU/mL, respectively. The correlation between ADNase B (y) and ASO (x) was y=0.4x+173 (r= 0.46). Mean ADNase B level (392 IU/mL) was significantly higher in children with S. pyogenes than in those with non-group A BHS (236 IU/mL) or no BHS (234 IU/ mL). Some schoolchildren were proven, via ASO and ADNase B tests, to be harboring asymptomatic S. pyogenes infections. The high ULN of ASO and ADNase B in schoolchildren should be carefully considered, in order to interpret the data collected from the patients. We could add the ADNase B test to our set of diagnostic tools, which would allow us to more accurately detect and diagnose streptococcal infections, as ADNase B was more specifically related to the results of throat cultures, and there was little correlation between ASO and ADNase B.
Antibodies, Bacterial/*blood ; Bacterial Proteins/immunology ; Child ; Deoxyribonucleases/*immunology ; Female ; Humans ; Korea ; Male ; Serologic Tests ; Streptococcal Infections/*diagnosis/*immunology ; Streptococcus pyogenes/enzymology/*immunology ; Streptolysins/immunology

OBJECTIVETo discuss the important role of actin polymerization in calcium ionophore A23187-induced human acrosome reaction and its mechanism.

METHODSEach spermatozoon specimen was divided into five groups, treated with A23187 3 micromol/L in Group A, Phalloidin 40 micromol/L and A23187 3 micromol/L in Group B, SLO 0.5 U/ml and A23187 3 micromol/L in Group C, SLO 0.5 U/ ml, Phalloidin 40 micromol/L and A23187 3 micromol/L in Group D, and nothing added in Grpup E. Then the percentage of the human acrosome reaction was assessed with Rhodamine-PSA (10 microg/ml).

RESULTSThe difference of the human spermatozoon acrosome reaction was significant (P < 0.01) among the 5 groups with or without SLO, Phalloidin and calcium ionophore A23187 but not between Groups A and B (P > 0.01).

CONCLUSIONPhalloidin does not work on the acrosome reaction of intact human spermatozoa, but in an SLO-permeabilized human spermatozoal model, it can obviously decrease the percentage of human spermatozoon acrosome reaction, which indicates that the polymerization of actin plays an important role in the course of human spermatozoon acrosome reaction, and mostly acts on the acrosome inside.

Acrosome Reaction ; drug effects ; Actins ; physiology ; Bacterial Proteins ; pharmacology ; Calcimycin ; pharmacology ; Cells, Cultured ; Humans ; Ionophores ; pharmacology ; Male ; Phalloidine ; pharmacology ; Spermatozoa ; drug effects ; physiology ; Streptolysins ; pharmacology