Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology最新文献

{"title":"The role of polymorphic short tandem (CA)n repeat loci segregation analysis in the detection of Duchenne muscular dystrophy carriers and prenatal diagnosis.","authors":"Veronica Ferreiro,&nbsp;Florencia Giliberto,&nbsp;Liliana Francipane,&nbsp;Irene Szijan","doi":"10.1007/BF03260074","DOIUrl":"https://doi.org/10.1007/BF03260074","url":null,"abstract":"<p><strong>Background: </strong>Duchenne and Becker muscular dystrophies (DMD/BMD) are X-linked diseases caused by mutations in the dystrophin gene at Xp21.2; they include gross deletions (60%), duplications (10%), and small mutations (30%). Since there is no cure or effective treatment for progressive muscular dystrophy, prevention of the disease is important and strongly depends on carrier-status information. Two-thirds of DMD/BMD cases are familial; thus, female relatives are candidates for carrier-risk assessment.</p><p><strong>Aim: </strong>Segregation analysis of polymorphic short tandem (CA)n repeats [STR-(CA)n] was used to establish and compare the haplotypes of female relatives of patients with DMD/BMD with those of the patient in order to identify the mutant dystrophin gene and thus determine each female relative's carrier status.</p><p><strong>Methods: </strong>248 individuals from 52 families were studied through segregation of up to 11 STR-(CA)n loci. The assay was performed on leukocyte DNA by PCR amplification, polyacrylamide-gel electrophoresis and autoradiography. Haplotypes were established by determination of alleles on the autoradiography.</p><p><strong>Results: </strong>38 of 51 (75%) female relatives from familial cases were diagnosed as carriers or non-carriers with a 95-100% likelihood, and 18 out of 56 (32%) female relatives from sporadic cases could be excluded from the risk of being a DMD carrier with the same probability. In addition, STR studies detected gross deletions in 13 of the 52 (25%) families in both male and female individuals, four of which were de novo deletions. STR assays were also informative in families without an available DNA sample of an affected male and in two of seven symptomatic females. Determination of carrier status was particularly significant for prediction of DMD risk in prenatal analysis of five male chorionic villi. Other genetic events revealed by STR analysis were: (i) 11 recombinations identified in 6.6% of meiosis in the DMD families; (ii) germinal mosaicism detected in two female carriers; and (iii) changes in STR-(CA)n length during transmission from father to daughters, including three retractions and one elongation at an estimated rate of 0.004.</p><p><strong>Conclusion: </strong>The STR assay is an excellent molecular tool for carrier-status identification and the detection of deletions and other genetic changes in families affected by DMD/BMD. Thus, it is useful in genetic counseling for the prevention of this disease.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 2","pages":"67-80"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF03260074","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25280879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"BCRABL transcript detection by quantitative real-time PCR : are correlated results possible from homebrew assays?","authors":"Sallyanne C Fossey,&nbsp;Andrea Ferreira-Gonzalez,&nbsp;Carleton T Garrett,&nbsp;Catherine I Dumur,&nbsp;Cindy L Vnencak-Jones","doi":"10.1007/BF03260090","DOIUrl":"https://doi.org/10.1007/BF03260090","url":null,"abstract":"<p><strong>Background: </strong>Quantitative real-time PCR has become the predominant molecular technique to monitor BCRABL levels in response to treatment in Ph(+) leukemia patients. However, without some form of standardized methodology between laboratories, the correlation of results is difficult.</p><p><strong>Methods: </strong>Using TaqMan-based assays, parallel quantitative real-time PCR analysis was performed on 70 clinical specimens at Vanderbilt University Medical Center and Virginia Commonwealth University. While the same positive control cell line (K562) and quality control gene (BCR) were used, the RNA isolation technique, cDNA synthesis, BCR control cell line, and PCR primer and probe sequences were different.</p><p><strong>Results: </strong>The detection of BCRABL-positive results spanned a dynamic range from 10(0) to 10(5)/100,000 cells. Forty-three samples were negative at both facilities. A Spearman rank correlation analysis was performed for the 22 BCRABL-positive paired results. The correlation coefficient, r(s), was 0.9435 (p < 0.00001), suggesting a strong correlation of the results. One discordant result was obtained for consecutive samples from one patient with a low BCRABL copy number as a result of a minimal RNA yield at one laboratory.</p><p><strong>Conclusions: </strong>These results suggest that quantitative real-time PCR assays for BCRABL detection can be comparable between laboratories despite significant differences in methodologies if the same positive control cell line and quality control gene are used. It is imperative that some level of assay standardization be adopted between laboratories, not only for patients who are monitored at different facilities, but also for larger investigative studies in which hematologic, cytogenetic and molecular responses are to be compared.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 4","pages":"187-93"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF03260090","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25784213","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Diagnostic proceeding in Silver-Russell syndrome.","authors":"Thomas Eggermann,&nbsp;Esther Meyer,&nbsp;Michael B Ranke,&nbsp;Martin Holder,&nbsp;Stefanie Spranger,&nbsp;Klaus Zerres,&nbsp;Hartmut A Wollmann","doi":"10.1007/BF03260093","DOIUrl":"https://doi.org/10.1007/BF03260093","url":null,"abstract":"<p><strong>Background: </strong>Silver-Russell syndrome (SRS) describes a uniform malformation syndrome characterized by pre- and postnatal growth restriction (<3rd percentile) and a typical craniofacial gestalt. The basic defect of SRS is currently unknown, and the number of meaningful genetic tests available is therefore limited. Different chromosomal aberrations have been identified, including in the chromosomal region 7p12-p14. Detailed analyses of numerous candidate genes have not revealed any relevant insights with respect to the etiology of the disease.However, maternal uniparental disomy (UPD) of chromosome 7 (matUPD7), the inheritance of both homologues of chromosome 7 only from the mother, is observed in approximately 10% of SRS patients. Here, we report on our experiences of UPD testing in patients referred to our laboratory with the clinical diagnosis of SRS. A diagnostic algorithm for SRS is suggested.</p><p><strong>Methods: </strong>Eighty-six patients with the clinical diagnosis of SRS were screened for matUPD7 by microsatellite typing. In 13 cases, the clinical data were consistent with the diagnosis of SRS. The other 73 patients were referred for UPD testing with the suspected diagnosis of SRS, but clinical data were scarce.</p><p><strong>Results: </strong>In total, we identified three new cases of matUPD7: one patient belonged to the cohort of 13 clinically characterized patients; the other two patients were referred with the suspected diagnosis of SRS but initially without detailed reports. DNA studies revealed uniparental heterodisomy 7 in two patients, while the results in the third case were consistent with uniparental isodisomy.</p><p><strong>Conclusions: </strong>MatUPD7 is predominantly detectable in patients showing SRS features, and testing should therefore be restricted to this group of growth-restricted patients. Generally, a combination of cytogenetic and molecular genetic tests can be offered in SRS, aiming at the detection of chromosomal rearrangements and matUPD7 in >10% of SRS patients.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 4","pages":"205-9"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF03260093","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25784216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Paraoxonase 1-192Q allele is a risk factor for idiopathic chronic pancreatitis.","authors":"Mariette Verlaan,&nbsp;Erik G A Harbers,&nbsp;Akos Pap,&nbsp;Jan B M J Jansen,&nbsp;Wilbert H M Peters,&nbsp;Joost P H Drenth","doi":"10.2165/00066982-200509010-00002","DOIUrl":"https://doi.org/10.2165/00066982-200509010-00002","url":null,"abstract":"<p><strong>Background: </strong>The cause of chronic pancreatitis (CP) remains unknown. However, oxidative stress might play a role since recent animal studies have demonstrated that oxygen-free radicals contribute to the pathogenesis of experimental pancreatitis. Human serum paraoxonase (PON1) is an antioxidant enzyme that protects against cellular damage from oxidative stress. Genetic variations resulting in variable activity rates of this enzyme, are of toxicological and physiological importance.</p><p><strong>Aim: </strong>We investigated whether genetic polymorphisms of the PON1 gene modify the risk for CP.</p><p><strong>Materials and methods: </strong>DNA samples were obtained from 236 adult CP patients of hereditary (n = 23), alcoholic (n = 137), or idiopathic (n = 76) origin. DNA from 113 healthy controls and from 93 alcoholic controls were analyzed for comparison. Patients and controls were all of Caucasian origin. Genetic polymorphisms (L55M and Q192R) in PON1 were determined by PCR, followed by restriction fragment length polymorphism analyses in all subjects.</p><p><strong>Results: </strong>The frequencies of the PON1-55 alleles did not differ between CP patients and healthy controls. However, the PON1-192Q allele was significantly more common in idiopathic CP patients (OR : 1.5, 95% CI 1.02, 2.5) compared with healthy controls.</p><p><strong>Conclusions: </strong>These data suggest that the PON1-192Q allele, resulting in partly deficient antioxidant and detoxification activity of this enzyme, might be a risk factor for idiopathic CP in Caucasians.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 1","pages":"9-15"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2165/00066982-200509010-00002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25204329","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Real-time Epstein-Barr virus PCR for the diagnosis of primary EBV infections and EBV reactivation.","authors":"Rianne Luderer,&nbsp;Marieke Kok,&nbsp;Hubert G M Niesters,&nbsp;Rob Schuurman,&nbsp;Okke de Weerdt,&nbsp;Steven F T Thijsen","doi":"10.1007/BF03260091","DOIUrl":"https://doi.org/10.1007/BF03260091","url":null,"abstract":"<p><strong>Background: </strong>The serological diagnosis of primary Epstein-Barr virus (EBV) infections is often difficult, whereas the relevance of elevated immunoglobulin G (IgG) antibodies against early antigen (EA) for the diagnosis of EBV reactivation has increasingly become a matter of dispute. Recently, EBV PCR has been added as a diagnostic tool. Positive EBV PCR has been demonstrated in the serum of patients with primary EBV infections and EBV reactivation.</p><p><strong>Objectives: </strong>To compare classical serological diagnosis of primary EBV infection and EBV reactivation with real-time EBV PCR.</p><p><strong>Study design: </strong>Sera from 45 patients were selected with detectable immunoglobulin M (IgM) antibodies against EBV viral capsid antigen (VCA), and 62 sera were selected with a reactivation profile. A real-time EBV PCR was performed with DNA extracted from these sera.</p><p><strong>Results: </strong>Based on serological data, the diagnosis of primary EBV infection was established for 24 of the 45 IgM VCA-positive patients. By performing PCR, seven extra cases of primary infection were diagnosed for which no heterophilic antibodies could be detected. In five cases of primary infection, no EBV DNA could be detected by PCR. Only in two of the 62 sera with a reactivation seroprofile could EBV DNA be detected.</p><p><strong>Conclusions: </strong>Based on these data, we suggest that for the diagnosis of primary infections, EBV PCR could lead to an increase of >16% in the number of positive diagnoses by confirming a positive IgM VCA in the absence of heterophilic antibodies. Furthermore, EBV PCR is positive in only 3% of sera with elevated antibodies against EA, raising doubt as to the utility of EA titers for diagnosing EBV reactivation.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 4","pages":"195-200"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF03260091","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25784214","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel mutation of the DHCR7 gene in a sicilian compound heterozygote with Smith-Lemli-Opitz Syndrome.","authors":"Fabrizio Romano,&nbsp;Barbara Fiore,&nbsp;Franca Maria Pezzino,&nbsp;Maria Teresa Longombardo,&nbsp;Angelo Baldassare Cefalù,&nbsp;Davide Noto,&nbsp;Ambra Puglisi,&nbsp;Alfio Brogna,&nbsp;Teresa Mattina,&nbsp;Maurizio Averna,&nbsp;Salvatore Travali","doi":"10.1007/BF03260092","DOIUrl":"https://doi.org/10.1007/BF03260092","url":null,"abstract":"<p><strong>Introduction: </strong>Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive disorder of cholesterol biosynthesis, resulting from deficient 7-dehydrocholesterol reductase (3beta-hydroxysterol Delta7-reductase) activity, the enzyme responsible for conversion of 7-dehydrocholesterol to cholesterol. SLOS is most common among people of European descent, with a reported incidence of 1 per 20,000-60,000 newborns, depending on the diagnostic criteria and the reference population. More than 80 different mutations have been identified in several hundred patients. In Italy, SLOS appears to be a rare condition, probably because of underdiagnosis.</p><p><strong>Method: </strong>We analyzed by direct sequencing the 7-dehydrocholesterol reductase gene (DHCR7) in a Sicilian patient with Smith-Lemli-Opitz syndrome and his parents in order to characterize the molecular defect.</p><p><strong>Results: </strong>The molecular analysis of the coding exons and the intron-exon boundaries of the DHCR7 gene demonstrated the presence of two missense mutations: a novel mutation (I251N) and a known mutation (E288K) responsible in a compound heterozygous state for a severe form of SLOS.</p><p><strong>Conclusion: </strong>The present study describes a Sicilian patient, a carrier of a novel mutation of the DHCR7 gene (I251N), which is responsible in a compound heterozygous state for a severe form of SLOS.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 4","pages":"201-4"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF03260092","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25784215","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Touch-down reverse transcriptase-PCR detection of IgV(H) rearrangement and Sybr-Green-based real-time RT-PCR quantitation of minimal residual disease in patients with chronic lymphocytic leukemia.","authors":"Sona Peková,&nbsp;Jana Marková,&nbsp;Petr Pajer,&nbsp;Michal Dvorák,&nbsp;Petr Cetkovský,&nbsp;Jirí Schwarz","doi":"10.2165/00066982-200509010-00004","DOIUrl":"https://doi.org/10.2165/00066982-200509010-00004","url":null,"abstract":"<p><strong>Background: </strong>Patients with chronic lymphocytic leukemia (CLL) can relapse even after aggressive therapy and autografts. It is commonly assumed that to prevent relapse the level of minimal residual disease (MRD) should be as low as possible. To evaluate MRD, highly sensitive quantitative assays are needed.</p><p><strong>Aim: </strong>The aim of the study was to develop a robust and sensitive method for detection of the clonal immunoglobulin heavy-chain variable (IgV(H)) rearrangement in CLL and to introduce a highly sensitive and specific methodology for MRD monitoring in patients with CLL who undergo intensive treatment.</p><p><strong>Methods: </strong>As a prerequisite for MRD detection, touch-down reverse transcriptase (RT)-PCR using degenerate primers were used for the diagnostic identification of (H) gene rearrangement(s). For quantitative MRD detection in 18 patients, we employed a real-time RT-PCR assay (RQ-PCR) making use of patient-specific primers and the cost-saving Sybr-Green reporter dye (SG). For precise calibration of RQ-PCR, patient-specific IgV(H) sequences were cloned.</p><p><strong>Results: </strong>Touch-down RT-PCR with degenerate primers allowed the successful detection of IgV(H) clonal rearrangement(s) in 252 of 257 (98.1%) diagnostic samples. Biallelic rearrangements were found in 27 of 252 (10.7%) cases. Degenerate primers used for the identification of clonal expansion at diagnosis were not sensitive enough for MRD detection. In contrast, our RQ-PCR assay using patient-specific primers and SG reached the sensitivity of 10(-)(6). We demonstrated MRD in each patient tested, including four of four patients in complete remission following autologous hematopoietic stem cell transplantation (HSCT) and three of three following allogeneic 'mini'-HSCT. Increments in MRD might herald relapse; aggressive chemotherapy could induce molecular remission.</p><p><strong>Conclusions: </strong>Our touch-down RT-PCR has higher efficiency to detect clonal IgV(H) rearrangements including the biallelic ones. MRD quantitation of IgV(H) expression using SG-based RQ-PCR represents a highly specific, sensitive, and economic alternative to the current quantitative methods.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 1","pages":"23-34"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2165/00066982-200509010-00004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25203599","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Use of duplex PCR-CTPP methods for CYP2E1RsaI/IL-2 T-330G and IL-1B C-31T/TNF-A T-1031C polymorphisms.","authors":"Yoshiko Atsuta,&nbsp;Haruya Kawase,&nbsp;Nobuyuki Hamajima,&nbsp;Kazuko Nishio,&nbsp;Yoshimitsu Niwa,&nbsp;Daisuke Tanaka,&nbsp;Kazuhito Yamamoto,&nbsp;Akiko Tamakoshi","doi":"10.1007/BF03260076","DOIUrl":"https://doi.org/10.1007/BF03260076","url":null,"abstract":"<p><strong>Background: </strong>Two duplex polymerase chain reaction (PCR) with confronting two-pair primer (PCR-CTPP) methods were designed for cytochrome P450 (CYP) 2E1 RsaI and interleukin (IL-2) T-330G, and for IL-1B C-31T and tumor necrosis factor-alpha (TNF-A) T-1031C. The four polymorphisms are considered to be functional, and the three cytokines reportedly inhibit CYP2E1 expression. Many studies have reported associations between the above polymorphisms and risk of diseases including cancers and inflammatory diseases.</p><p><strong>Aim: </strong>The main objective of this study was to examine the applicability of the established PCR conditions to a real situation.</p><p><strong>Participants: </strong>Participants were female examinees aged from 35 to 85 years who attended health checks run by a local government in Japan.</p><p><strong>Results: </strong>The allele frequencies among 325 female health check examinees were 0.804 for CYP2E1 c1 allele, 0.668 for IL-2-330T allele, 0.554 for IL-1B-31T allele, and 0.822 for TNF-A-1031T allele. p-Values from a Hardy-Weinberg equilibrium test were 0.658, 0.955, 0.062, and 0.806, respectively.</p><p><strong>Discussion: </strong>Clear DNA bands observed with electrophoresis allowed us to genotype the four polymorphisms. The genotype frequencies were within the Hardy-Weinberg equilibrium test proportions, though the p-value for IL-1B C-31T was marginal.</p><p><strong>Conclusions: </strong>Both duplex PCR-CTPP methods may be useful tools for studies on the association between these polymorphisms and disease risk.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 2","pages":"89-94"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF03260076","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25280883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Heart-fatty acid-binding protein as a marker for early detection of acute myocardial infarction and stroke.","authors":"Pierre Lescuyer,&nbsp;Laure Allard,&nbsp;Denis F Hochstrasser,&nbsp;Jean-Charles Sanchez","doi":"10.2165/00066982-200509010-00001","DOIUrl":"https://doi.org/10.2165/00066982-200509010-00001","url":null,"abstract":"<p><p>Heart-fatty acid-binding protein (H-FABP) is a small cytosolic protein involved in intracellular fatty acid transport. This protein, highly concentrated in the heart, is quickly released into plasma after myocardial injury. Results from numerous studies suggest that H-FABP is an excellent marker for the early detection of myocardial damage. H-FABP is also expressed in the brain, although in lower concentrations than in the heart. Recent preliminary studies also investigated the usefulness of H-FABP for the diagnosis of acute and chronic neurological disorders. These potential applications of H-FABP in clinical practice are reviewed in this article, with a strong focus on the early diagnosis of acute myocardial infarction and stroke.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 1","pages":"1-7"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2165/00066982-200509010-00001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25204328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Protein chip for detection of different HCV antibodies: preparation, quality control, and clinical evaluation.","authors":"Wen Zhang,&nbsp;Jian Huang,&nbsp;Mei-Fen Zhou,&nbsp;Li-Yan Chen,&nbsp;Ya-Ping Ding,&nbsp;Heng-Jie Cao,&nbsp;Yong-Yao Geng,&nbsp;Sheng-Qi Wang","doi":"10.1007/BF03260075","DOIUrl":"https://doi.org/10.1007/BF03260075","url":null,"abstract":"<p><strong>Introduction: </strong>As a contagious disease caused by hepatitis C virus (HCV) hepatitis C is a serious threat to human health. Therefore, the detection and verification of HCV infection is very important in the treatment of hepatitis C. This study investigated the preparation, quality control, and clinical evaluation of a protein chip capable of simultaneously detecting different HCV antibodies. The aim was to establish a convenient method for the detection of HCV.</p><p><strong>Method: </strong>To prepare the protein chip, six antigens including five recombinant HCV antigens (chimeric, core, NS3, NS4, and NS5) and interleukin (IL)-1 were arrayed onto aldehyde-coated slides and blocked using 10% calf serum in phosphate buffered saline. After dilution with sample solution, the serum sample was added to a reaction well on the protein chip. After incubation for 30 minutes at 37 degrees C, fluorescence Cy3-labeled rabbit antihuman IgG was added and incubated again for 30 minutes at 37 degrees C, and then scanned. Positive or negative controls were established from serum samples with or without HCV infection. Clinical evaluation was done by detecting 490 serum samples using the protein chips and ELISA reagents, with 150 of the 490 serum samples confirmed by recombinant immunoblot assay (RIBA).</p><p><strong>Results: </strong>The protein chip for detection of five HCV antibodies was successfully prepared. Fifteen positive controls and 15 negative controls were established as standard samples for quality control. The quality control-passed protein chip was tested again using the standard of the National Institute for the Control of Pharmaceutical and Biological Products (NICPBP), and met the quality control criteria prescribed by the NICPBP. In the clinical evaluation with 490 samples, the coincidence rates between the protein-chip assay and ELISA were 97.4% for positive and 100% for negative results. Five inconsistent samples that were positive in ELISA, but non-positive (four samples) or negative (one) in the protein-chip assay, were confirmed by RIBA (gold standard) to be four non-positive and one negative. The results of 150 samples showed the coincidence rates between protein chip and RIBA were 98.15% for positive and 96.88% for single-segment positive.</p><p><strong>Conclusion: </strong>The protein-chip assay has higher sensitivity and specificity than ELISA and has a high coincidence rate with RIBA. The protein chip, characterized by its easy operation and low economic cost, will be very useful for in vitro detection of HCV antibodies.</p>","PeriodicalId":79690,"journal":{"name":"Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology","volume":"9 2","pages":"81-7"},"PeriodicalIF":0.0,"publicationDate":"2005-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/BF03260075","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25280881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}