摘要:

AbstractThe sensitivity and application of the polymerase chain reaction (PCR) for the diagnosis of parvovirus B19 (B19) infection was investigated by simultaneously assaying a collection of 279 consecutively received samples for presence of anti‐B19 IgM and IgG antibodies by Western blot and for B19 DNA by PCR and dot‐blot hybridization (dot‐blot); samples were sera from patients with suspected B19 infection. PCR and dot‐blot detected B19 DNA in 9% (16/179) and 1% (2/179), respectively of Ab‐positive samples (IgM+/IgG‐,IgM+IgG+,IgM‐IgG+), and in 28% (15/54) and 2% (1/54), respectively, of IgM+ samples. PCR also detected B19 DNA in 2% (2/100) of IgM‐/IgG‐ samples, both of which had normal total IgG and IgM levels. PCR is of unique value because it permits diagnosis of B19 infection even in the absence of specific acute phase (IgM) and in the presence or absence of convalescentphase (IgG) Ab. © 19

ISSN:0887-8013    

DOI:10.1002/jcla.1860060402

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractA microparticle‐enhanced nephelometric immunoassay was developed for myoglobin quantitation in human serum. It uses rabbit antimyoglobin serum and hydrophilic polyacrylic microparticles covalently coated with baboon myoglobin in a competitive immunoagglutination system. The level of microparticle agglutination is assessed with a specially designed nephelometer. This sensitive (45 ng/ml of myoglobin detected in serum) and accurate (coefficients of variation from 3.0% to 8.2% in precision study and linear recovery of myoglobin in overloaded sera) immunoassay was evaluated with human sera from patients suffering from acute myocardial infarction. Myoglobin levels in patient's serum on admission appeared to be correlated with clinical and biological parameters assessed in emergency wards and later. This new, rapid, and easy microparticle‐enhanced nephelometric immunoassay could thus be useful in emergency conditions for the early quantitation of serum myoglo

ISSN:0887-8013    

DOI:10.1002/jcla.1860060403

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractWe describe an enzyme‐linked immunosorbent assay (ELISA) to measure apolipoproteins Al and B secreted by Hep G2 cells and in cell homogenates. These assays utilize commercially available polyclonal antibodies, affinity‐purified to improve their specificity, thereby achieving a dramatic increase in the sensitivity of the assay. These affinity‐purified antibodies were also more sensitive than a series of monoclonal antibodies tested. We achieved a sensitivity of 0.4 ng in the apo Al assay, and a sensitivity of 5 ng in the apo B assay. By these methods, we measured secretion rates by Hep G2 cells of 358 ± 41 ng/mg cell protein/hr for apo B and 137 ± 8 ng/mg cell protein/hr for apo Al. These assays also allowed the measurement of intracellular apolipoproteins and thus can be used to facilitate investigations of human lipoprotein metabolism in cell culture systems. © 1992 Wiley

ISSN:0887-8013    

DOI:10.1002/jcla.1860060404

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractA simple and sensitive procedure for total mercury in whole blood and urine using inductively coupled plasma‐mass spectrometry (ICP‐MS) is described. Specimens are prepared by precipitation‐extraction with 50% v/v hydrochloric acid containing EDTA and cysteine, centrifuged, and filtered through fritended screening column; the filtrates are directly analyzed by ICP‐MS. The method is linear between 2 and 200 μg/L in the specimen with an absolute sensitivity of 0.2 μg/L in the final supernatant. The assay variability at various concentrations (μg/L) of mercury are as follows: intra‐assay whole blood (n = 20)—4.6 ± 0.6(c.v. 12.3%), 18.3 ± 1.1(c.v.6.1%), 56.4 ± 2.8(c.v.5.0%); inter‐assay whole blood (n = 15)—5.7 ± 1.0(c.v.16.8%), 19.7 ± 2.7(c.v. 13.5%), and 50.1 ± 6.9(c.v.13.7%); urine (n = 20)—9.3 ± 1.2 (c.v. 12.9%), 29.6 ± 2.2 (c.v. 7.4%). Recovery of organic and inorganic mercury from blood samples ranges from 91.6% to 110.2%. The method is suitable for analysis of total mercury, both organic and inorganic, in whole blood and u

ISSN:0887-8013    

DOI:10.1002/jcla.1860060405

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThe effect of acid pH treatment of circulating immune complexes (CIC) derived from tuberculosis sera was studied on the relative titers of specific antibody (CIC Ab) and mycobacterial antigen (CIC Ag) in the complexes. While the specific antibody titers increased, the titers of CIC Ag declined as a result of acid pH treatment of CIC, both changes being highly significant statistically. Direct exposure of TB sera to pH 2.8 also resulted in significant enhancements in the titers of antibodies directed againstMycobacterium tuberculosis(M.tb.). Competition experiments indicated that the acid pH‐treated antibodies retained their antigen specificity. TB sera treated with pH 2.8 for 30 min and the neutralized back to pH 7.4 retained the enhanced antibody reactivity even after 7 days of storage at 4°C. Our results indicate that acid pH treatment induces higher antigen binding ability in anti‐M.tb. antibodies, present in free or complexed form in TB sera. These changes appear to be irreversible. Dissociation of CIC and analyzing the titers of released antibody and antigen components offered no advantage with respect to the immunodiagnosis of tuberculosis, as compared to the levels of undissociated CIC components or the serum antibody. © 1992 Wiley‐Li

ISSN:0887-8013    

DOI:10.1002/jcla.1860060406

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractA novel and ultrasensitive noncompetitive enzyme immunoassay (hetero‐two‐site complex transfer enzyme immunoassay) for α‐human atrial natriuretic peptide (α‐hANP) in plasma is described. α‐hANP was biotinylated using N‐hydroxysuccinimidobiotin and trapped onto an anti‐α‐hANP [6–28] IgG‐coated polystyrene ball. After washing, biotinylated α‐hANP was eluted from the polystyrene ball with HCI and was reacted with 2,4‐dinitrophenyl‐fluorescein‐bovine serum albumin‐disulfide‐rabbit anti‐α‐hANP [6–28]IgG conjugate. The complex formed was trapped onto (anti‐2,4‐dinitrophenyl group) IgG‐coated polystyrene balls and, after washing, reacted with avidin‐β‐D‐galactosidase conjugate. The polystyrene balls were washed, and the complex of the three components was eluted with ϵN‐2, 4‐dinitrophenyl‐L‐lysine and transferred to anti‐fluorescein IgG‐coated polystyrene balls. After washing, the complex was released from the polystyrene balls by reduction with 2‐mercaptoethylamine and transferred to (anti‐rabbit IgG) IgG‐coated polystyrene balls. β‐D‐Galactosidase activity bound to the last polystyrene balls was assayed by fluorometry. The detection limit of α‐hANP [1–28] was 3 fg (1 amol)/tube. Interference by plasma proteins was eliminated by separation of peptides from proteins using a molecular sieve. The assay range of plasma α‐hANP [1–28]was 0.04–120 ng/L, and plasma levels of hANP

ISSN:0887-8013    

DOI:10.1002/jcla.1860060407

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractWe have compared the size, the binding to Concanavalin A (Con A), and the affinity for monoclonal antibody 1116NS‐199 (Mab 19‐9) among CA 19‐9 molecules from sera of cystic fibrosis (CF) and pancreatic carcinoma patients and from sputum extracts. CA 19‐9 molecules of two different sizes were found in all types of specimens by Sepharose 4B chromatography. While the smaller CA 19‐9 molecule was predominant in CF patient sera, the larger molecule was associated with most of the sera from patients with pancreatic carcinoma. The majority of the sputum extracts contained the larger CA 19‐9 molecule. All CA 19‐9 molecules studied by Con A chromatography did not appear to bind to Con A, and almost 100% were found in the nonreactive fraction. The CA 19‐9 molecules from sera of either CF or pancreatic carcinoma patients exhibited variable affinities for Mab 19‐9, some approaching that of the standard curve but many also having lower affinities. The lowest affinity was displayed by CA 19‐9 molecules from the sputum extract. It appears that development of more specific assays for CF and for carcinoma is possible if the correct CA 19‐9 molecule is selected for antibody preparation and for use as standards.

ISSN:0887-8013    

DOI:10.1002/jcla.1860060408

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractEight commercially available enzyme‐linked immunoadsorbent assays (ELISA) for the detection of cytomegalovirus (CMV)‐specific IgM were used in parallel to determine the presence of CMB‐IgM in 123 serum samples from pregnant women. The results obtained with the eight kits were compared. Based on concordance of six or more of the eight kits, we assessed sensitivity, specificity, and overall agreement, as well as incidence of false‐positive and ‐negative results for each kit. The results obtained by ELISA were then compared with those obtained by immunofluorescence (IF) and immunoblotting (IB).Our study did not single out one outstanding ELISA kit among the eight evaluated, nor did it suggest that IF or IB are better than ELISA. Furthermore our results indicate that IB might be useful in several cases as, beside its good sensitivity, most IB‐false‐positive sera are easily recognized as reacting exclusively with pp 150, the unique reactivity to pp 150 not being among the IB profiles of IB‐true‐positive sera. Nevertheless 14.6% of sera remained CMV‐IgM‐indeterminate.

ISSN:0887-8013    

DOI:10.1002/jcla.1860060409

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractSerum amyloid A (SAA) and C‐reactive protein (CRP) levels were compared in 830 serum samples from 155 cystic fibrosis (CF) patients. Correlation coefficients were calculated for all samples (r=0.73), for samples from non‐corticosteroid treated (CFNS) patients (n=698, r=0.80), and for samples from corticosteroid treated (CFS) patients (n=132, r=0.35). SAA was the more sensitive indicator of pulmonary inflammation when SAA and CRP were compared to pulmonary function tests of 49 hospitalized patients at admission and discharge. CRP levels were significantly (p<.05) lower at admission in CFS patients than in CFNS patients, whereas SAA levels were not significantly different between the two groups. All nine CFS patients hospitalized had elevated SAA levels (average 22 times above normal limits) at admission, while only six had elevated CRP levels (average 3.7 times above normal limits) at admission. In the 40 CFNS patients both SAA and CRP levels were significantly elevated at admission. In each case SAA and CRP levels declined as pulmonary functions improved with effective antimicrobial therapy. In three instances SAA levels increased during hospitalization while CRP levels did not. In each case, rising SAA levels indicated clinical deterioration associated with evolving resistance ofP. aeruginosawhich required a change in antibiotic therapy. © 1992 Wiley‐Lis

ISSN:0887-8013    

DOI:10.1002/jcla.1860060410

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThe specificity of a tumor marker (CA54/61) and its individual epitopes (CA54 and CA61) recognized by monoclonal antibodies (MA54 and MA61) and expressed by the same tumor marker were studied. Serum levels of CA54 and CA61 were compared with that of CA54/61. In lung adenocarcinoma and ovarian carcinoma, the positive rates of CA61 (42% and 68%) were higher than those of CA54 (32% and 32%) and similar to those of CA54/61 (45% and 74%). The serum levels of CA54 and CA61 showed a significant correlation (r=0.78), but 22% of tested sera were positive for CA54 and negative for CA61 or negative for CA54 and positive for CA61. It was demonstrated that the tumor specificity between CA54 and CA61 was not same and that the tumor specificity of CA54/61 was similar to that of CA61 rather than CA54. Moreover, the difference in the tumor specificity between CA54 and CA61 was considered to be reflected in the difference in their epitope structure. © 1992 Wiley‐Liss, I

ISSN:0887-8013    

DOI:10.1002/jcla.1860060411

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY