• Parasites, Hosts and Diseases
• /
• v.25 no.2
• /
• pp.159-167
• /
• 1987

To analyse the antigen specificity of patients sera from 24 confirmed neurocysticercosis and a monoclonal antibody, SDS-PAGE using 10~15% linear gradient gel and EITB were done. Cystic fluid, saline extracts of scolex and of whole worm of C. cellulosae, saline extracts of sparganum, hydatid cyst fluid, saline extracts of Fasciola, Clonorchis and Paragonimus were used as antigen. Of protein bands in cystic fluid of C. cellulosae, patient sera reacted frequently to bands of 152, 94, 64, 48, 24, 15, 10 and 7kDa proteins. To saline extracts of scolex and whole worm of C. cellulosae, patients sera reacted frequently to 94, 64, 52, 39, 34, 15 and 10kDa bands. Two bands in sparganum extract (130 and 64kDa) and two bands in hydatid cyst fluid (52 and 27kDa) were cross-reacting bands with sera from cysticercosis patients. Saline extracts of Fasciola, ClonorchiJ and Paragonimus did 'not exhibit cross-reacting bands. Monoclonal antibody to cystic fluid of C. cellulosae was found to react with low molecular weight proteins of 15, 10 and 7kDa.

• Annals of Clinical Microbiology
• /
• v.18 no.2
• /
• pp.37-43
• /
• 2015

Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.

• Parasites, Hosts and Diseases
• /
• v.24 no.1
• /
• pp.25-41
• /
• 1986

The applicability of micro-ELISA was evaluated in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of $2.5{\;}{\mu}g/ml$. Serum was diluted to 1 : 100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of o. 18 in both samples. The results were summarized as follows: 1. The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1% ; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. 2. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5%; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. 3. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 ot 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. 4. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. 5. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes.

• Parasites, Hosts and Diseases
• /
• v.26 no.2
• /
• pp.73-86
• /
• 1988

The sensitivity and specificity of crude and affinity-purified antigens of Clcnorchis sinensis obtained from the infected rabbits were studied. Stage-specific antigenic proteins from the eggs, metacercariae and adult worms were characterized by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent astray (ELISA). The results were as follows: 1. The antibody.binding antigen (ABA) purified from whole worm crude antigen (IVWA) by CNBr-activated Sepharose 4B affinity chromatography made :l specific bands against rabbit antisera on Ouchterlony gel diffusion plate, while WWA made 7 bands. Major WWA protein bands by SDS-PAGE were found at 16, 300~18, 500 and 28, 000~29, 000 daltons, while major ABA protein bands were at 18, 000~21, 000 and 29, 000~31, 000 daltons. The reactivity of ABA with rabbit anti-sera in ELISA was remarkably less sensitive than that of WWA. 2. Molecular weights of egg antigen (EGA), metacercarial antigen (MEA) and adult worm antigen (WWA) of C. sinensis ranged from 15, 000-200, 000 daltons, 15, 000-100, 000 daltons and 11, 000~80, 000 daltons, respectively. Major WWA proteins consisted mainly of polypeptide bands of low molecular weight, less than 31, 000 daltons, while those of EGA and MEA consisted of higher molecular T.eights than 30, 000 daltons. 3. The ELISA reactivities of WWA to rabbit anti.sera were remarkably greater than those of MEA. EGA showed negative reaction throughout the experiments. WWA showed higher optical density (O.D.) than 1.0, when reacted with rabbit anti-sera obtained at 4~6 weeks after the infection. In the rabbit anti-sera later than 12 weeks after the infection, the O.D. reacting witll WWA showed a plateau without variation. MEA shoT.ed relatively low O.D. values (<0.6), when reacted with anti-sera from lightly in(ected groups throughout the experiments, althougll there were some wealth positive cases (O.D.>0.6) ill heavily infected groups. MEA reacted with rabbit anti-sera showed negative results on Ouchterlony gel diffusion plates. Summarizing the above results, it is suggested that the whole worm antigen prepared from the adult worms of C. sinensis is most highly antigenic. However, this antigen might reveal cross reactions with other trematodes such as Paragonimus westermani, therefore, purification of antigenic proteins from the crude antigen is essential 18 increase the sensitivity and specificity for the immuncdiagnosis of clonorchiasis.

• Journal of Life Science
• /
• v.22 no.8
• /
• pp.1114-1119
• /
• 2012

Bacteria are frequently contaminated during the collection and processing procedures of boar semen. Of the contaminants, Stenotrophomonas (S.) maltophilia is a Gram-negative bacterium that is widely distributed in a variety of habitats. Although PCR assays have been developed for the detection of S. maltophilia, they cross-react with some species of Xanthomonas. In this study, we designed a primer set for the detection of S. maltophilia in order to target the chiA (GenBank accession no. NC_010943) gene. The specific PCR products were amplified from S. maltophilia only, not from other tested strains that are frequently found in semen. The detection limit of the PCR was $1.5{\times}10^3$ CFU/ml with pure-cultured S. maltophilia and $1.5{\times}10^4$ CFU/ml with S. maltophilia spiked in semen. Twenty-six (5.9%) S. maltophilia were isolated from 440 semen samples. The PCR results exhibited 98.9% agreement with a comparison of S. maltophilia isolation. Also, the sensitivity and specificity of the PCR were 100% and 98.7%, respectively. In the antimicrobial susceptibility test, S. maltophilia isolates were highly susceptible to enrofloxacin and florfenicol, while the majority of them were resistant to amoxicillin/clavulanic acid, apramycin, ceftiofur, penicillin, and spectinomycin. These results indicated that the PCR using the chiA gene was proven to be reliable and effective for the detection of S. maltophilia with high levels of sensitivity and specificity.

• Applied Biological Chemistry
• /
• v.15 no.2
• /
• pp.111-142
• /
• 1972

An experimental Chung-Kook-Jang was prepared using the strain Bacillus subtilis sp. isolated by the author Samples were taken in 12 hrs interval during the fermentation and the oligopeptides were separated by the method of molecular sieving using the ion exchange resin column of Dowex-50. Only the X-16 fraction of oligopeptides was taken and the components of oligopeptides were developed in two dimensional thin layer chromatograms. The each peptide spot was eluted and each peptide was isolated. The pattern and kinds of amino acids, and N and C-terminal amino acids were studied. Fourteen different oligopeptides could be detected by the two dimensional thin layer chromatography, all of which were consisted of $4{\sim}9$ kinds of amino acids. No dipeptides and no tripeptides could be found. The N and C-terminal amino acids and the residual component amino acids of all these 14 peptides could be summarized as the follows. [P]-I. Pro (Cys Ala Asp Trp Ile Val) Glu [P]-II. Val (His Arg Glu Thr Ala Met) Asp [P]-III. Glu (Cys Lys Asp Thr Met) Ala [P]-IV. Glu(His Ser Ala) Met) [P]-V. Ile (Cys Asp Arg Gly Pro T.p Phe) His [P]-VI. Gly(Asp ser) Lys [P]-VII. Thr(Pro Tyr Phe) Asp [P]-VIII. Phe(Tyr Leu Ile) Val [P]-IX. Trp (Phelle) Thr [P]-X. Ile (Arg Leu) Phe [P]-XI. Asp (Lys His Ser Gly Glu Pro) Ala [P]-XII. Glu (Cys Asp Gly) Ser [P]-XIII. Ala (Arg Tyr) Glu [P]-XIV. Met (Glu Ala) His It appears that the protease of the Bacillus subtilis K-27 syrain has rather wider range of specificity than proteases of Aspergoillus soya, pepsin, chymotrypsin, and trypsin.

• The Korean Journal of Mycology
• /
• v.17 no.2
• /
• pp.82-90
• /
• 1989

Detection of antibody against pathogenic fungi in serum specimens of the patients with pulmonary tuberculosis or other lung diseases has been carried out(male) using the indirect fluorescence antibody technique and immunodiffusion tests. Immunodiffusion tests revealed that 104(36.5%) out of 285 patients examined showed a positive precipitin reaction against one or more of fungal antigens. The majority of ID positive patients 64(61.5%) reacted with Aspergillus fumigatus antigen and 49(47.1%) patients reacted with Candida albicans antigen ID positive reaction to A. fumigatus was found little more frequently among male patients, while Candida albicans reactors were found more frequently among female patients. Age distribution of ID positive reactors was high(49.1-43.3%) in age group of 40-59 years, but least or none in age group of less than 30 years. Age of fungal mycelium used as antigen did not effect sensitivity of the indirect flubrescence (IF) technique in detecting antibody to A. fumigatus. Antibody class against A. fumigatus that showed highest titer was IgG and thus FITC labeled anti-IgG immunoglobulin shoul be preferable. As relatively large amount of cell wall components of Aspergilli shared antigenically, a considerable cross-reaction was observed among A. fumigatus, A. flavus and A. niger, but not much with C. albicans. While (IF) has much better sensitivity when compared with ID, relative specificity of the latter procedure cannot to be overried, so that they could be batter used together in order to obtain quantitative measurement of antibody with relative specificity.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
• /
• v.12 no.4
• /
• pp.359-369
• /
• 2007

Amoebophrya is an obligate endoparasitic eukaryotic dinoflagellate infecting host species and eventually killing them within a short period. Because of its host specificity and significant impacts on population dynamics of host species, it has long been proposed to be a potential biological agent for controlling harmful algal bloom (HAB). For several decades, the difficulties of culturing host - parasite systems have been a great obstacle to further research on the biology of Amoebophrya but recent success of several culture systems reactivates this research field. In this study, as a preliminary work for understanding the impacts of Amoebophrya on the population dynamics of host species, semimonthly occurrence of infected host dinoflagellates by Amoebophrya spp. had been observed in Jinhae Bay for two years and with a host - parasite system cultivated, host specificity of Amoebophrya spp. on several dinoflagellates was tested. Amoebophrya spp. were observed in the cellular organelle and cytoplasm of several species including Akashiwo sanguinea, Ceratium fusus, Dinophysis acuminata, Heterocapsa triquetra, Oblea sp., Prorocentrum minimum, P. triestinum, Scrippsiella spinifera, and S. trochoidea. Among them two host - parasite systems for an athecate dinoflagellate, A. sanguinea, and for a thecate dinoflagellate, H. triquetra, had been able to be successfully established as laboratary cultures. Cross-infection tests for 6 species of dinoflagellates in which Amoebophrya was observed or had been reported to exist confirmed high preference for host species of the parasite. Through the continuous research on Amoebophrya occurring in Korean coastal waters, we need to maintain various host - parasite culture systems, which will be very helpful for understanding its ecological role in marine food webs and for applying the species to biologically control harmful algal blooms.

• Journal of Pharmaceutical Investigation
• /
• v.38 no.4
• /
• pp.261-267
• /
• 2008

The aim of the present study was to evaluate the bioequivalence of two gabapentin preparations. We used Neurontin tablet 800 mg (Pfizer Korea Inc.) as a reference drug for bioequivalence of Gabalep tablet 800 mg (Chong Kun Dang Pharmaceutical Co., Korea), and performed this whole study according to the guidelines of Korea Food and Drug Administration (KFDA). Twenty five healthy male volunteers were administered with each drug in a randomized $2{\times}2$ cross-over study with one week washout interval. After drug administration, blood was taken at predetermined time intervals ($0{\sim}24$ hours) and the concentrations of gabapentin in serum were determined using an high performance liquid chromatography-tandem mass spectrometer (LC-MS/MS) employing electrospray ionization technique and operating in multiple reaction mornitoring (MRM). The analytical method was validated in specificity, accuracy, precision and linearity. The phar-macokinetic parameters such as AUCt and Cmax were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed AUCt and Cmax. $Mean{\pm}SD$. of AUCt and Cmax value for reference drug and test drug were $29.94{\pm}9.23\;({\mu}g/mL{\cdot}hr)$ and $3.12{\pm}1.11\;({\mu}g/mL{\cdot}hr)$, and $31.48{\pm}9.77\;({\mu}g/mL{\cdot}hr)$ and $3.15{\pm}1.03\;({\mu}g/mL)$, respectively. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log(0.8) to log(1.25) for AUCt and Cmax, respectively. These results indicate that Gabalep tablet 800 mg is bioequivalent to Neurontin tablet 800 mg.

• IMMUNE NETWORK
• /
• v.2 no.3
• /
• pp.175-181
• /
• 2002

Background: S100A6 is a calcium-binding protein overexpressed in several tumor cell lines including melanoma with high metastatic activity and involved in various cellular processes such as cell division and differentiation. To detect S100A6 protein in patient' samples (ex, blood or tissue), it is essential to produce a monoclonal antibody specific to the protein. Methods: First, cDNA coding for ORF region of human S100A6 gene was amplified and cloned into the expression vector for GST fusion protein. We have produced recombinant S100A6 protein and subsequently, monoclonal antibodies to the protein. The specificity of anti-S100A6 monoclonal antibody was confirmed using recombinant S100A recombinant proteins of other S100A family (GST-S100A1, GST-S100A2 and GST-S100A4) and the cell lysates of several human cell lines. Also, to identify the specific recognition site of the monoclonal antibody, we have performed the immunoblot analysis with serially deleted S100A6 recombinant proteins. Results: GST-S100A6 recombinant protein was induced and purified. And then S100A6 protein excluding GST protein was obtained and monoclonal antibody to the protein was produced. Monoclonal antibody (K02C12-1; patent number, 330311) has no cross-reaction to several other S100 family proteins. It appears that anti-S100A6 monoclonal antibody reacts with the region containing the amino acid sequence from 46 to 61 of S100A6 protein. Conclusion: These data suggest that anti-S100A6 monoclonal antibody produced can be very useful in development of diagnostic system for S100A6 protein.