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1.
J Investig Clin Dent ; 10(4): e12473, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31631564

ABSTRACT

AIM: The aim of the present study was to correlate the immunoexpression of alpha-smooth muscle actin (α-SMA) for myofibroblasts with the serum levels of transforming growth factor-ß1 (TGF-ß1) in oral submucous fibrosis (OSMF). METHODS: A total of 100 cases of histopathologically confirmed OSMF were assessed for α-SMA expression. Clinical data, such as age, sex, mouth opening, and habit history, were obtained for each case. Serum TGF-ß1 levels were recorded in 73 patients with the help of enzymelinked immunosorbent assay technique. RESULTS: The staining index of α-SMA increased concomitantly with higher myofibroblast count in the increasing histopathological grades of OSMF (P ≤ .05). Serum TGF-ß1 levels were highest in the intermediate grades of OSMF. Clinical parameters, such as mouth opening, cheek flexibility, and tongue protrusion, showed a direct correlation with increasing clinical grades of OSMF. CONCLUSIONS: The progressive increase in myofibroblasts from early to advanced stages suggests their potential use as markers for evaluating the severity of OSMF. Additionally, as myofibroblasts are responsible for producing a variety of factors that are involved in the fibrotic processes; they could be the key link in the pathogenesis of OSMF. Interruption of their development, recruitment, or activation could provide a unique therapeutic target for future treatment options in patients with OSMF.


Subject(s)
Oral Submucous Fibrosis , Actins , Humans , Muscle, Smooth , Transforming Growth Factor beta1 , Transforming Growth Factors
2.
J Immunoassay Immunochem ; 40(4): 407-418, 2019.
Article in English | MEDLINE | ID: mdl-31088248

ABSTRACT

Derris scandens (Roxb.) Benth. is a medicinal plant used for treatment of musculoskeletal pain in Thai traditional medicines. Its stem contains active compound genistein-7-O-[α-rhamnopyranosyl-(1 to 6)-ß-glucopyranoside] (GTG) which is used as a biomarker for standardization of D. scandens extracts. As an alternative for rapid quantitation of GTG, a monoclonal antibody against GTG was prepared and applied for an indirect competitive enzyme-linked immunosorbent assay (ELISA) to determine GTG in plants and herbal products. The established method provided a quantification range of 0.31-10 µg/mL with a limit of detection of 0.29 µg/mL. The assay was validated for precision and accuracy by intra- and interassay variation analyses, recovery test, and comparison analysis between the amounts of GTG determined by ELISA and HPLC. The results exhibited that the developed ELISA is sensitive and effective for determination of GTG in D. scandens plant materials and herbal products.


Subject(s)
Antibodies, Monoclonal/immunology , Derris/chemistry , Enzyme-Linked Immunosorbent Assay/methods , Genistein/analysis , Plant Extracts/analysis , Plant Extracts/immunology , Quality Control , Chromatography, High Pressure Liquid , Genistein/analogs & derivatives , Genistein/immunology
3.
Rev Sci Tech ; 38(3): 761-786, 2019 12.
Article in English, French | MEDLINE | ID: mdl-32286568

ABSTRACT

In Algeria, the prevalence of causes of abortion on dairy cattle farms (whether infectious causes or not) has been little studied. The current study involved a serological analysis conducted between October 2014 and June 2016 in northern Algeria using an enzyme-linked immunosorbent assay test on blood samples taken from 368 cows that had aborted on 124 farms. It was complemented by a survey to identify the factors associated with a higher or lower risk of exposure to Coxiella burnetii, Chlamydia abortus and Toxoplasma gondii, using univariate logistic regression and then multivariate logistic regression. The individual serological prevalences obtained were 8.4% (31/368) for C. burnetii and 12.2% (45/368) for C. abortus. For T. gondii, the individual seroprevalence was 13.8% (51/368); the factors associated with a higher risk of individual exposure were the fourth month of gestation (odds ratio [OR] = 22.68; 95% confidence interval [CI]: 1.38-392.97) and the fifth month of gestation (OR = 25.51; 95% CI: 1.47-442.11). All the other factors identified by the multivariate logistic regression were associated with a lower risk of exposure. They are the inspection visits in 2015 (OR = 0.0006; 95% CI: 0.000004-0.12) and in 2016 (OR = 0.0005; 95% CI: 0.000002-0.13) and artificial insemination (OR = 0.15; 95% CI: 0.05-0.44) for C. burnetii ; winter (OR = 0.39; 95% CI: 0.15-1.00), spring (OR = 0.45; 95% CI: 0.20-0.97), and artificial insemination (OR = 0.27; 95% CI: 0.13-0.56) for C. abortus; and the number of gestations (OR = 0.38; 95% CI: 0.16-0.92) for T. gondii. The seroprevalence at herd level was 16.1% (20/124) for C. burnetii and 29.8% (37/124) for both C. abortus and T. gondii. At herd level, the risk factors associated with a higher risk of exposure to C. abortus and T. gondii were the practice of deworming (OR = 3.89; 95% CI: 1.53-9.89) and drilling individual wells as a source of drinking water (OR = 7.50; 95% CI: 2.11-26.69). For C. burnetii, the inspection visit in 2015 (OR = 0.02; 95% CI: 0.0008-0.65) and in 2016 (OR = 0.01; 95% CI: 0.0003-0.36), artificial insemination (OR = 0.21; 95% CI: 0.06-0.69) and rodent eradication (OR = 0.19; 95% CI: 0.06-0.57) were factors that reduced the risk of exposure.


En Algérie, la prévalence des causes d'avortement dans les élevages bovins laitiers (que ces causes soient d'origine infectieuse ou non infectieuse) a été peu étudiée. Cette étude concerne une analyse sérologique conduite d'octobre 2014 à juin 2016 dans le nord de l'Algérie à l'aide d'un test ELISA (méthode immuno-enzymatique) sur des prélèvements sanguins issus de 368 vaches ayant avorté provenant de 124 élevages et complétée par un formulaire d'enquête visant à identifier, par une régression logistique uni puis multivariée, les facteurs associés à un risque augmenté ou diminué d'exposition à Coxiella burnetii, Chlamydia abortus et Toxoplasma gondii. Les prévalences sérologiques individuelles obtenues ont été respectivement de 8,4 % (31/368) pour C. burnetii et de 12,2 % (45/368) pour C. abortus. Pour T. gondii, la séroprévalence individuelle était de 13,8 % (51/368), avec comme facteurs associés à un risque accru d'exposition individuelle, le quatrième mois de gestation (rapport des cotes [odds ratio : OR] = 22,68 ; intervalle de confiance [IC] à 95 % : 1,38-392,97) et le cinquième mois de gestation (OR = 25,51 ; IC à 95 % : 1,47-442,11). Les autres facteurs identifiés par la régression logistique multivariée étaient tous des facteurs associés à un risque diminué d'exposition. Ils concernaient l'année de visite en 2015 (OR = 0,0006 ; IC à 95 % : 0,000004-0,12) et en 2016 (OR = 0,0005 ; IC à 95 % : 0,000002-0,13) et l'insémination artificielle (OR = 0,15 ; IC à 95 % : 0,05-0,44) pour C. burnetii ; l'hiver (OR = 0,39 ; IC à 95 % : 0,15-1,00), le printemps (OR = 0,45 ; IC à 95 % : 0,20-0,97) et l'insémination artificielle (OR = 0,27 ; IC à 95 % : 0,13-0,56) pour C. abortus ; et le nombre de gestations (OR = 0,38 ; IC à 95 % : 0,16-0,92) pour T. gondii. La séroprévalence obtenue à l'échelle du troupeau a été respectivement de 16,1 % (20/124) pour C. burnetii et de 29,8 % (37/124) pour C. abortus et T. gondii. À l'échelle des troupeaux, les facteurs associés à un risque accru d'exposition à C. abortus et T. gondii concernent respectivement la pratique du déparasitage (OR = 3,89 ; IC à 95 % : 1,53-9,89) et le forage personnel comme source d'abreuvement (OR = 7,50 ; IC À 95 % : 2,11-26,69). Pour C. burnetii, l'année de visite en 2015 (OR = 0,02 ; IC à 95 % : 0,0008-0,65) et en 2016 (OR = 0,01 ; IC à 95 % : 0,0003-0,36), l'insémination artificielle (OR = 0,21 ; IC à 95 % : 0,06-0,69) et l'éradication des rongeurs (OR = 0,19 ; IC à 95 % : 0,06-0,57) sont des facteurs de diminution du risque d'exposition.


En Argelia está poco estudiada la prevalencia de las causas de aborto en las explotaciones bovinas de producción láctea (ya sean causas de tipo infeccioso o no infeccioso). Los autores exponen aquí un análisis serológico realizado entre octubre de 2014 y junio de 2016 en el norte de Argelia. Empleando una técnica de ensayo inmunoenzimático (ELISA), se examinaron muestras sanguíneas de 368 vacas que habían sufrido un aborto procedentes de 124 explotaciones, análisis que se complementó con un formulario de investigación destinado a determinar, por regresión logística de una sola variable y después multifactorial, los factores ligados a un aumento y a un decremento del riesgo de exposición a Coxiella burnetii, Chlamydia abortus y Toxoplasma gondii. Las prevalencias serológicas individuales obtenidas fueron, respectivamente, de un 8,4% (31/368) para C. burnetii y un 12,2% (45/368) para C. abortus. Por lo que respecta a T. gondii, se obtuvo una seroprevalencia individual del 13,8% (51/368). Los factores ligados a un mayor riesgo de exposición individual eran el cuarto mes de gestación (razón de probabilidades [odds ratio: OR] = 22,68; intervalo de confianza [IC] al 95%: 1,38-392,97) y el quinto mes de gestación (OR = 25,51; IC al 95%: 1,47-442,11). Todos los otros factores determinados por regresión logística multifactorial venían asociados a una reducción del riesgo de exposición. Se trataba concretamente de: el hecho de que la visita se hubiera efectuado en 2015 (OR = 0,0006; IC al 95%: 0,000004-0,12) o en 2016 (OR = 0,0005; IC al 95%: 0,000002-0,13) y la inseminación artificial (OR = 0,15; IC al 95%: 0,05-0,44), en el caso de C. burnetii; el hecho de que fuera invierno (OR = 0,39; IC al 95%: 0,15-1,00) o primavera (OR = 0,45; IC al 95%: 0,20-0,97) y la inseminación artificial (OR = 0,27; IC al 95%: 0,13-0,56), en el caso de C. abortus; y el número de gestaciones (OR = 0,38; IC al 95%: 0,16-0,92), en el caso de T. gondii. La seroprevalencia de rebaño obtenida fue respectivamente de un 16,1% (20/124) para C. burnetii y de un 29,8% (37/124) para C. abortus y T. gondii. A la escala de los rebaños, los factores vinculados a un mayor riesgo de exposición a C. abortus y T. gondii eran, respectivamente, la práctica de desparasitaciones (OR = 3,89; IC al 95%: 1,53-9,89) y el uso de un pozo propio como abrevadero (OR = 7,50; IC al 95%: 2,11-26,69). En cuanto a C. burnetii, los factores que reducían el riesgo de exposición del rebaño eran el hecho de que el año de visita fuera 2015 (OR = 0,02; IC al 95%: 0,0008-0,65) o 2016 (OR = 0,01; IC al 95%: 0,0003-0,36), la inseminación artificial (OR = 0,21; IC al 95%: 0,06-0,69) y la erradicación de los roedores (OR = 0,19; IC al 95%: 0,06-0,57).


Subject(s)
Abortion, Veterinary/microbiology , Abortion, Veterinary/parasitology , Cattle Diseases/epidemiology , Chlamydia Infections/veterinary , Q Fever/veterinary , Toxoplasmosis, Animal/epidemiology , Algeria/epidemiology , Animals , Cattle , Cattle Diseases/microbiology , Cattle Diseases/parasitology , Chlamydia , Chlamydia Infections/epidemiology , Coxiella burnetii , Dairying , Enzyme-Linked Immunosorbent Assay , Female , Pregnancy , Prevalence , Q Fever/epidemiology , Risk Factors , Seroepidemiologic Studies , Toxoplasma
4.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-841707

ABSTRACT

Objective: To compare the urinary neutrophil gelatinase-associated lipocalin (NGAL) levels in the acute kidney injury (AKI) and non-AKI premature infants, and to explore the clinical value of NGAL level in the early diagnosis of AKI. Methods: A total of 85 premature infants were divided into AKI group (n=30) and non-AKI group (n=55) according to whether AKI occurred. The levels of urinary NGAL were detected by enzymelinked immunosorbent assay (ELISA) method at 24, 48, 72, 120, and 168 h after birth. The levels of serum creatinine (Scr) of preterm infants were detected by dry chemical method. The receiver operation characteristics (ROC) curve is used to judge the diagnostic value of each indicator. Results: There were no significant differences in gender, gestational ages, body weights at birth, ages at NICU of the premature infants between two groups (P> 0. 05). There were significant differences in the urinary NGAL levels of the premature infants between two groups at different time points (F=62. 710, P<0. 01); the urinary NGAL level was increased at 24 h after birth, reached the highest peak at 72 h afger birth, and decreased from 72 h after birth. The urinary NGAL levels of the premature infants in AKI group were higher than those in non-AKI group at 24, 48, 72 and 120 h after birth (P< 0. 05). There were statistically significant differences in the Scr levels of the pretmature infants between two groups at different time points (F=27. 332, P<0. 05). The Scr levels of the premature infants in AKI group were higher than those in non-AKI group at 48, 72 and 120 h after birth (P<0. 05). The area under ROC curve (AUC) of urinary NGAL level at 24 after birth was significantly greater than that of Scr level (P<0. 01). When the cutoff value of urinary NGAL level was 6. 95 ng middot; dL-1, the sensitivity and specificity of urinary NGAL level in diagnosis of AKI were 0. 900 and 0. 836, respectively. Conclusion: The level of urinary NGAL can effectively predict the AKI in the premature infants.

5.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-841934

ABSTRACT

Objective: To explore the differences in the positive rates of specific-IgE (sIgE) of the common allergens between the cow's milk protein allergy (CMPA) and healthy infants and the distribution characteristics of positive sIgE of common allergens in the CMPA infants, and to provide basis for comprehensive intervention of the CMPA infants. Methods: A total of 156 cases of CMPA and 318 cases of healthy infants were selected as the subjects. The serum sIgE and total IgE levels of common allergens of the infants in two groups were detected by enzyme-lmked immunosorbent assay. The differences in the positive rates of serum sIgE of common allergens and total IgE of the infants in two groups were compared. Results: There were no statistical differences in the positive rates of serum sIgE and total IgE for common allergens of the infants between CMPA group and healthy group (P> 0.05). The major food allergens with high positive rates of sIgE in CMPA group were cow's milk 44.2%), egg white (10.3%) and cashew nut (5.1%), and the inhale allergens with high positive rates of sIgE were cat hair 21.2%), dog hair (9.6%) and house dust mite 4.5%). While for the healthy infants, the major food allergens with high positive rates of sIgE were cow's milk 45%), egg white (14.2%) and cashew nut (6.0%), and the inhale allergens with high positive rates of sIgE were cat hair (25.8%), dog hair (14.5%) and fungus combinations 4.5%). The analysis in different age groups (0.05). Conclusion: There are no significant differences in the serum sIgE and total IgE positive rates between the CMPA and healthy infants. The detection of serum sIgE and total IgE of common allergens is of little clinical significance for the CMPA infants.

6.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-691578

ABSTRACT

Objective:To explore the differences in the positive rates of specific-IgE(sIgE)of the common allergens between the cow's milk protein allergy(CMPA)and healthy infants and the distribution characteristics of positive sIgE of common allergens in the CMPA infants,and to provide basis for comprehensive intervention of the CMPA infants.Methods:A total of 156 cases of CMPA and 318 cases of healthy infants were selected as the subjects.The serum sIgE and total IgE levels of common allergens of the infants in two groups were detected by enzyme-linked immunosorbent assay.The differences in the positive rates of serum sIgE of common allergens and total IgE of the infants in two groups were compared.Results:There were no statistical differences in the positive rates of serum sIgE and total IgE for common allergens of the infants between CMPA group and healthy group(P>0.05).The major food allergens with high positive rates of sIgE in CMPA group were cow's milk(44.2%),egg white(10.3%)and cashew nut(5.1%),and the inhale allergens with high positive rates of sIgE were cat hair (21.2%),dog hair(9.6%)and house dust mite(4.5%).While for the healthy infants,the major food allergens with high positive rates of sIgE were cow's milk(45%),egg white(14.2%)and cashew nut(6.0%),and the inhale allergens with high positive rates of sIgE were cat hair(25.8%),dog hair(14.5%)and fungus combinations(4.5%).The analysis in different age groups(<1 year old and 1-2 years old)showed that there were no statistical differences in the positive rates of serum sIgE and total IgE for common allergens of the infants between CMPA group and healthy group(P>0.05).Conclusion:There are no significant differences in the serum sIgE and total IgE positive rates between the CMPA and healthy infants.The detection of serum sIgE and total IgE of common allergens is of little clinical significance for the CMPA infants.

7.
Vet Immunol Immunopathol ; 183: 1-6, 2017 Jan.
Article in English | MEDLINE | ID: mdl-28063471

ABSTRACT

West Nile virus (WNV) mainly infects birds, horses and humans. Outcomes of the infection range from mild uncharacteristic signs to fatal neurologic disease. The main objectives of the present study were to measure serum IgG and IgM antibodies in naturally exposed and vaccinated horses and to compare results of haemagglutination inhibition test (HIT), enzyme-linked immunosorbent assay (ELISA) and plaque reduction neutralisation test (PRNT). Altogether 224 animals were tested by HIT for WNV antibodies and 41 horses were simultaneously examined by ELISA and PRNT. After primary screening for WNV antibodies, horses were vaccinated. Samples were taken immediately before and 3-5 weeks after each vaccination. McNemar's chi-squared and percent agreement tests were used to detect concordance between HIT, ELISA and PRNT. Analyses by HIT confirmed the presence of WNV antibodies in 27/105 (26%) naturally exposed horses. Sera from 57/66 (86%) vaccinated animals were positive before the first booster and from 11/11 (100%) before the second booster. HIT was less sensitive for detecting IgG antibodies. We could detect postvaccination IgM in 13 cases with IgM antibody capture ELISA (MAC-ELISA) and in 7 cases with HIT. WNV is endemic in Hungary and regularly causes natural infections. Protective antibodies could not be measured in some of the cases 12 months after primary vaccinations; protection is more reliable after the first yearly booster. Based on our findings it was not possible to differentiate infected from recently vaccinated horses using MAC-ELISA. HIT cannot be used as a substitute for ELISA or PRNT when detecting IgG, but it proved to be a useful tool in this study to gain statistical information about the tendencies within a fixed population of horses.


Subject(s)
Antibodies, Viral/blood , Enzyme-Linked Immunosorbent Assay/veterinary , Hemagglutination Inhibition Tests/veterinary , Horse Diseases/virology , Viral Vaccines/immunology , West Nile Fever/veterinary , West Nile virus , Animals , Horse Diseases/blood , Horse Diseases/immunology , Horses , West Nile Fever/immunology
8.
Cancer Research and Clinic ; (6): 472-474, 2012.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-429168

ABSTRACT

ObjectiveTo study the value of serum phosphopyruvate hydratase PH protein for the diagnosis of cerebral injuries in patients with brain malignant tumor.MethodsSerum PH protein levels were detected by enzyme-linked immunosorbent assay in 56 patients with brain malignant tumor. Compare the differences among the status of cerebral injuries.And compare the differences among the patients with general with different radiotherapy methods 32 cases,three dimensional conformal radiothrapy 24 cases,and different peritumoral brain edema levels(cmild 18 cases, moderate ao cases severe 30 cases). Results Before radiotherapy the levels of serum PH protein in patients and the health control were (4.12±0.56),(4.66±0.62)μg/L,no significant differece(P>0.05).And there was also no significant difference between the before and after radiotherapy for the cerebroma,the levels were(7.84±0.72) μg/L,(t=3.89,P=0.001 ).The PH levels of general radiation therapy and three dimensional comformal radiotherapy were (13.59±0.92),(6.14±0.52)μg/L.There was cignificant difference(P=0.002) After radiotherapy,The levels of serum PH protein of the different dropsical degree,mild,moderate and severe were(4.47:±0.55),(6.17±0.62),(15.21±0.86) μg/L, respectively,showing significant difference(F=15.61,P=0.0001).The therapies influenced the serum PH levels(P<0.05)ConclusionHigh levels of serum PH protein are associated with severe cerebral injuries in brain malignant tumor.So high serum PH level may serve as an progressive predictor of the injury.

9.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-420051

ABSTRACT

Objective To establish a biotin-streptavidin ELISA method to measure CTGF,and evaluate the clinical value of CTGF for the diagnosis of liver fibrosis in chronic hepatitis B (CHB) patients.Methods Biotinylated anti-CTGF polyclonal antibody and monoclonal antibodies were prepared for the establishment of this biotin-streptavidin ELISA method.Two hundreds and sixty-four CHB patients were subjected into non-or mild liver fibrosis group (S0-S1,108 cases) and severe liver fibrosis group (S2-S4,156 cases),according to the liver biopsy pathological diagnosis.The CTGF assay's diagnostic capacity for CHB was assessed by comparing the area under ROC (AUC) with that of a panel of hepatic fibrosis markers ( HA,PC Ⅲ,C Ⅳ,LN and APRI).Analysis of variance and rank sum test were performed to carry out comparisons between multiple groups.Student's t-test and Mann-Whitney U test were performed for the pairwise comparison between multiple samples.Spearman rank correlation test was performed to analyze the correlation between different hepatic fibrosis stages.Results The minimum detectable dose and detection rang of the ELISA was 0.2 μg/L and 0-64 μg/L respectively.The intra-assay and inter-assay CV at high and low level were 4.5%,9.8% and 10.1%,12.8% respectively.Serum CTGF concentrations in S0-S1 group and S2-S4 group were 6.7(3.1 - 10.1 ) μg/L and 16.1 ( 11.8 -27.2) μg/L,with a statistically significant difference (U =1 217,P <0.001 ).There was a significant correlation between the levels of serum CTGF and fibrosis stages ( r =0.689,P < 0.001 ),AUC of CTGF was 0.841 (95% CI:0.762 - 0.920) in distinguishing mild fibrosis from significant fibrosis.When the cut-off value of CTGF was 10.3 μg/L,the sensitivity and specification was 70.5% and 82.4% respectively.The sensitivity of parallel combination test of CTGF and APRI was 96.1%,which was higher than that of HA (75.6%),PC Ⅲ (70.5% ),C Ⅳ(63.6%),LN(79.5% ),APRI(86.3% ).The specificity of the combination test was 65.5%,which was lower than of above liver fibrosis markers [HA ( 72.5% ),PC Ⅲ ( 76.5% ),C Ⅳ ( 78.4% )].The specificity of serial combination test of CTGF and PC Ⅲ was 95.9%,which was higher than that of HA,PC Ⅲ,CⅣ,LN(64.7% ),APRI(66.1% ),however,the sensitivity of the combination test was 67.7%,which was lower than that of above HA,PC Ⅲ,and APRI.Conclusions The biotin-streptavidin ELISA method measuring serum CTGF has a high minimu detectable dose sensitivity,and specificity.Serum CTGF level is significantly correlated with fibrosis stage,and CTGF maybe a valuable marker for liver fibrosis assessment.The paralledl combination of CTGF and APRI could be used as screening for significant liver fibrosis markers.The serial combination of CTGF and PC Ⅲ may be considered as a confirmatory diagnostic marker for liver fibrosis.

10.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-380809

ABSTRACT

Objective To clone and express the human asialoglycoprotein receptor(ASGPR) H1 subunit, purify and identify the immunoreactivity of the recombinant protein, and establish the enzyme linked immunosorbent assay (ELISA) to detect anti-ASGPR antibodies in diagnosis of autoimmune hepatitis. Methods The CRDHI cDNA (435 bp) was subcloned into eukaryotic vector PEGH, and the recombinant protein expression was induced by D (+)-Galactose. The recombinant CRDH1 was purified with Glutathione Sepharose 4B, and its immunoreactivity was identified by SDS-PAGE and western blot as well as MALDI-TOF. ELISA was established to detect the anti-ASGPR antibodies in serum samples of 45 patients with AIH, 30 patients with SLE, 30 patients with RA, 10 patients with SS and 30 normal controls. Results The sequencing of recombinant plasmid showed the CRDH1 gene was successfully inserted to the eukaryotic expression vector with correct sequence and open reading frame. The fusion protein showed a molecular weight of 42 500 Da on SDS-PAGE gel and confirmed to be the human ASGPR by MALDI-MS through peptide mass fingerprint analysis with Mascot in human protein database. It shared 98. 34% homology with ASGPR H1 subunit. Western blot analysis showed that the fusion protein had the same immunoreactivity as human ASGPR. The results of ELISA indicated that the positive rate of anti-ASGPR was 35.6% ( 16/45 ), but the ELISA was negative in other control. There was significant difference of positivity of the autoantibodies between AIH and non-AIH controls (χ2 = 31.85,P < 0. 01 ). Conclusions The human plasmid containing ASGPR is successfully clone into Saccharomyces cerevisiae Y258. The recombinant autoantigen owns good antigenicity and specificity. ELISA established with the purified protein shows good specificity for diagnosis of AIH.

11.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-683290

ABSTRACT

Objective To study clinical significance of anti-momoner C-reactive protein (anti- mCRP) antibody in systemic lupus erythematosus (SLE) and assess the relationship between serum CRP and anti-mCRP antibody.Methods Enzyme-linked immunosorbent assay (ELISA) was applied to determine serum level of anti-mCRP antibody in 113 pateints with SLE,65 patients with other rheumatic diseases,including primary Sjgren syndrome,rheumatoid arthritis,osteoarthritis,ankylosing spondylitis and systemic sclerosis,and 32 healthy controls.Serum level of CRP was evaluated by turbidimetry.Clinical manifestations and laboratory indicators of the patients were all recorded.Results Serum level of anti- mCRP antibody in SLE patients was significantly higher than that in patients with other rheumatic diseases and healthy controls,respectively (t=2.502 and 5.352,respectively,P 0.05).Titer of anti-mCRP antibody closely correlated with systemic lupus erythematosus disease activity index score (r=0.248,P0.05). Conclusions Level of Anti-mCRP antibody increased significantly in patients with SLE,which associated with disease activity of SLE and can be used as a valuable marker in evaluating activity of SLE.

12.
Yonsei Medical Journal ; : 84-90, 2001.
Article in English | WPRIM (Western Pacific) | ID: wpr-147205

ABSTRACT

Malaria is one of the most important parasitic diseases especially in tropical areas. Over 300 million people are affected and the condition causes 1-3 million deaths each year. It is transmitted by the bite of infected Anopheles mosquitoes. Although Korea was declared to be free of Malaria by the WHO in 1979, malaria re-emergence has been apparent since 1993 amongst soldiers located near the De-Militarized Zone (DMZ) in the northern part of the country. Conventional microscopic examination of thin and thick blood films demonstrates the presence of the parasite and thus this method has been used to confirm the diagnosis of malaria, but it is a labor-intensive procedure and relies upon subjective interpretation. To overcome these limitations, fast and reliable methods for malaria detection have been recently introduced. In this study, we compared three kinds of antibody detection kits and one biochemical test kit that determines the presence of Plasmodium lactate dehydrogenase (pLDH) with conventional peripheral blood smears. The antibody detection methods examined were, two rapid test pack format methods and a single microplate format enzyme-linked immunosorbent assay (ELISA) kit, as manufactured by Korean companies. The sensitivities of the three commercial antibody detection kits in the early stage of malaria were 70.8%, 77.4%, and 63.6%, their corresponding specificities 90.5%, 91.8%, and 80.9%, and their accuracies 87.6%, 87.0%, and 76.7%. The sensitivity and specificity of the pLDH assay were 100% apiece and the results were in 100% concordance with the microscopy of thick blood films. Thus, the pLDH assay may be used as an alternative for conventional microscopic blood film examination, especially in emergency situations when prompt treatment is necessary.


Subject(s)
Adult , Aged , Female , Humans , Male , Animals , Antibodies, Protozoan/blood , Enzyme-Linked Immunosorbent Assay , L-Lactate Dehydrogenase/blood , Malaria/diagnosis , Middle Aged , Plasmodium/enzymology , Sensitivity and Specificity
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