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At least 973 records · Page 54Linked to original sources
PubMed2003

Detection of serum anti-ganglioside antibodies by latex agglutination assay in Guillain-Barré syndrome: comparison with ELISA.

OBJECTIVE: Rapid detection of serum anti-ganglioside antibodies in Guillain-Barré syndrome (GBS) could facilitate early diagnosis and early initiation of treatment, which might shorten the term of illness and reduce sequelae. We examined serum anti-ganglioside antibodies in patients with GBS using the latex agglutination assay developed by Alaedini and Latov (J Immunoassay 21: 377-386, 2000) with some modifications. MATERIALS AND METHODS: We used 75 sera from GBS patients, which exhibited IgG anti-GM1, GD1b, or GQ1b, or IgM anti-GM2 antibodies on previous enzyme-linked immunosorbent assay (ELISA). Blue latex beads (2.5% solution of 0.3 microm) were coated with 1 mg/ml of GM1, GD1b, GQ1b or GM2. Aliquots (4 microl) of serum and the ganglioside-coated particles were mixed and rocked on a glass slide for 30 to 40 seconds. The reaction was observed under a microscope and compared with the antiganglioside antibody titers determined with ELISA. RESULTS: Agglutination was strong in sera of which the IgM or IgG titers of anti-GM1, GD1b, GQ1b or GM2 antibodies were found to be more than 1:6,400 on ELISA except for 2 samples, but weak or absent in sera with titers of 1:3,200. Agglutination was absent in sera of which the antibody titers were less than 1:3,200 on ELISA. CONCLUSION: We could rapidly detect serum IgM and IgG anti-GM1, GD1b, GQ1b and GM2 antibodies in patients with GBS by means of the latex agglutination assay when sera exhibited high titers of the respective antibodies on ELISA. The sensitivity of our agglutination assay was much lower than that of ELISA.

Antibodies, Anti-Idiotypic↗
PubMed2006

[Detection of bcr/abl fusion gene from K562 cell line and mononuclear cells of CML patients by DNA-PCR].

OBJECTIVE: To detect bcr/abl fusion gene at DNA level in K562 cell line and mononuclear cells from patients with chronic myelogenous leukemia (CML). METHODS: Based on previous research, a set of DNA-PCR primers was redesigned. DNA from K562 cells and mononuclear cells of CML patients was extracted respectively. After DNA-PCR for bcr/abl fusion gene the amplified fragments were then sequenced. RESULT: At DNA level the bcr/abl fusion gene in K562 cells and mononuclear cells of 2 CML patients was amplified. Furthermore, the DNA breakpoint of fusion gene in the above samples through sequencing of amplified fragments was localized. CONCLUSION: DNA-PCR offers a new detection technology for bcr/abl fusion without the expression of fusion gene.

Fusion Proteins, bcr-abl↗
PubMed2000

[Detection of epidermal growth factor receptor gene amplification in human laryngeal carcinomas by means of fluorescence in situ hybridization].

OBJECTIVE: To detect epidermal growth factor receptor(EGFR) gene amplification in human laryngeal carcinoma Hep-2 cell line and laryngeal carcinoma tissues. METHODS: The technique of fluorescence in situ hybridization(FISH). RESULTS: The EGFR gene amplification of a laryngeal carcinoma Hep-2 cell line and 5 laryngeal carcinoma tissues were detected by FISH. In the metaphase chromosome and interphase nuclei of Hep-2 cell line and 2 laryngeal carcinoma tissues, distinct cluster and multiple dot signals were found. In the interphase nuclei of the other 3 laryngeal carcinoma tissues, no increase in the number or extent of the hybrid signals was found. CONCLUSION: Compared with normal diploid cell, the EGFR gene amplification was observed at different levels ranging from 2 to 8 folds in metaphase chromosome and interphase nuclei of Hep-2 cell line and 2 laryngeal carcinoma tissues while no amplification was observed in the other 3 laryngeal carcinoma tissues. The results demonstrate that quantitative detection of amplified gene by FISH in the metaphase chromosome and interphase nuclei is useful for detection of laryngeal carcinoma tissues.

ErbB Receptors↗
PubMed2001

Carrier detection in non-deletional Duchenne/Becker muscular dystrophy families using polymorphic dinucleotide (CA) repeat loci of dystrophin gene.

BACKGROUND & OBJECTIVES: Carrier detection and prenatal diagnosis is of great importance for families with one or more sons affected with Duchenne/Becker muscular dystrophy (D/BMD). In about 35-40 per cent of these patients, the causative mutation does not involve gross rearrangement in the structure of dystrophin gene. In these non-deletional families, genetic counselling can be provided only by linkage analysis. The aim of the present study was to determine the carrier status of female relatives in north Indian families with non-deletional D/BMD using highly polymorphic intragenic dinucleotide (CA) repeat markers. METHODS: Six short tandem repeats (STRs) spanning 5' (1), central (4) and 3' regions of the dystrophin gene were used to analyse 14 unrelated families comprising 68 individuals with 12 female siblings at risk of being carriers. RESULTS: Five female siblings inherited at risk STR haplotype, six inherited normal haplotype and one had meiotic recombination. The intragenic recombinations were observed in three families at the central region STR loci and in one family between the proximal and central regions of the gene. INTERPRETATION & CONCLUSIONS: Our study suggested that at least 6 STR markers spanning 5', central and 3' regions of the dystrophin gene are essential to ascertain one or more informative loci and to rule out recombinations in non-deletional D/BMD families for carrier analysis.

Dinucleotide Repeats↗
PubMed2002

Detection of occult metastases in lymph nodes from patients with colorectal carcinoma by reverse transcriptase-polymerase chain reaction.

OBJECTIVE: To detect occult metastases in lymph nodes from patients with colorectal carcinoma and its clinical significance. METHODS: The metastases in 260 lymph nodes from 39 histologically verified colorectal cancer patients were studied by both hematoxylin-eosin (HE) staining and cytokeratin-20 (CK20) specific RT-PCR. Ten normal lymph nodes were served as negative controls, and HT29 colon cancer cell line and 5 colorectal cancer specimens as positive controls. RESULTS: Ten normal lymph nodes were CK20-negative, HT29 cells and 5 tumor specimens were all CK20-positive. All 29 lymph nodes from 16 patients which confirmed metastases by HE staining exhibited CK20 positive expression; an additional 28 lymph nodes from 5 patients with no histologically detectable metastases expressed CK20 mRNA, i.e. presence of metastases. The difference of the positivity was significant (11.1% vs 21.9%, P < 0.01). According to the HE staining, the cases of Dukes' A, B, C and D were 3, 20, 12 and 4, respectively. In the 20 patients of Dukes' B stage, 5 of them had CK20-positive lymph nodes. CONCLUSION: CK20-specific RT-PCR is a highly sensitive, specific and simple method for detecting occult metastases in lymph nodes. The detection of CK20 mRNA expression in lymph nodes is recommended to precisely determine tumor stage and postoperative adjuvant therapy for patients with colorectal cancer, and further studies should be done in future to confirm the findings.

Adult↗
PubMed2005

[Perioperative detection of MAGE-1mRNA and AFP mRNA in peripheral blood of patients with hepatocellular carcinoma: relationships with recurrence].

OBJECTIVE: To detect MAGE-1 mRNA and AFP mRNA in the peripheral blood of patients with hepatocellular carcinoma (HCC) perioperatively, and explore their relationships with recurrence. METHODS: Peripheral blood from 45 patients with HCC, obtained perioperatively, was tested for the presence of MAGE-1 mRNA and AFP mRNA by nested RT-PCR. All patients were followed for metastatic recurrence for average 11 months after surgery. The peripheral blood from 22 patients with hepatitis B and cirrhosis, 11 patients with hepatic hemangioma, 12 patients with metastatic liver cancer and 20 healthy volunteers served as control. RESULTS: Four of the twelve (33.3%) metastatic hepatic cancer patients were positive for MAGE-1 mRNA, and three of the twenty-two (13.6%) patients with hepatitis B and cirrhosis were positive for AFP mRNA while the other controls kept negative in their peripheral blood. Detection rates for MAGE-1 mRNA/AFP mRNA in the peripheral blood of patients with HCC were 42.2%/51.1% before operation, 20%/24.4% seven days after surgery, and 15.6%/22.2% 28 days later respectively. In chi-square analysis, detection of peripheral-blood MAGE-1 mRNA and AFP mRNA 28 days after surgery indicated metastatic recurrence. Furthermore, the two-marker detection had a higher specificity for prediction of metastatic recurrence than single marker detection. CONCLUSION: Combined detection of cancer-specific MAGE-1 mRNA and hepatocyte-specific AFP mRNA by nested RT-PCR in the peripheral blood of patients with HCC 28 days after surgery may be useful for the prediction of metastatic recurrence.

Adult↗
PubMed2006

[Detection of CD95 and CD95L mRNA expression after liver transplantation using SYBR real-time PCR during acute rejection].

OBJECTIVE: To detect the expression of CD95 and CD95L mRNAs after liver transplantation and investigate the role of CD95 and CD95L in acute liver allograft rejection. METHODS: The expressions of CD95 and CD95L mRNAs of peripheral blood lymphocyte from 56 liver allograft recipients were examined using SYBR real-time PCR. RESULTS: CD95 and CD95L mRNA levels in the recipients with acute rejection were significantly higher than those without rejection (P<0.01), and the elevation occurred about 2 days earlier than that of alanine aminotransferase and aspartate aminotransferase. CONCLUSION: CD95 and CD95L are related to acute liver allograft rejection and their mRNA expression level may serve as an indicator for prediction and diagnosis of acute rejection episodes.

Acute Disease↗
PubMed1997

Detection of antibodies to Aspergillus fumigatus in serum of horses with mycosis of the auditory tube diverticulum (guttural pouch).

OBJECTIVE: To detect antibodies against Aspergillus fumigatus antigens in serum samples from horses and to evaluate the relevance of this method as an alternative approach to the diagnosis of mycosis of the auditory tube diverticulum (guttural pouch mycosis [GPM]). ANIMALS: Twelve clinically normal horses (controls) and 12 horses with GPM diagnosed by endoscopic observation of characteristic mycotic plaques. PROCEDURE: Antibodies to A fumigatus antigens were detected in serum by use of an ELISA and immunoblot analysis with extracellular antigens. RESULTS: Antibodies against A fumigatus antigens were found in healthy and diseased horses. Titer of total Aspergillus antibodies was not diagnostic for GPM. In contrast, immunoblot analysis results indicated that 2 antigens of 22 and 26 kd were constantly recognized by sera from diseased horses. CONCLUSIONS: Reactivity to 22- and 26-kd A fumigatus antigens, as measured by immunoblot analysis, seemed to be diagnostic for GPM in horses.

Animals↗
PubMed2000

Transmitral Doppler: a new transthoracic contrast method for patent foramen ovale detection and quantification.

OBJECTIVES: This study compared a new transthoracic echocardiographic (TTE) method for detection of right to left bubble passage, transmitral Doppler (TMD), against two-dimensional (2D) TTE contrast study and the gold standard, of transesophageal echocardiography (TEE), and assessed its utility in quantitative assessment of patent foramen ovales (PFO). BACKGROUND: Current TTE methods are relatively insensitive in PFO detection and do not allow quantitative assessment of right to left shunt. METHODS: In 44 patients (59 years, range 34 to 76 years) saline contrast and color Doppler studies were performed in three conditions--TTE TMD, TTE 2D and TEE. Bubble transit on the TMD was measured semiquantitatively by a visual bubble score and objectively by integrating the acoustic power within the mitral velocity envelope. RESULTS: By TEE it was determined that 17 patients (39%) had PFOs; 16 had right to left contrast passage, and only 1 had left to right flow by color Doppler. Against TEE contrast study, the sensitivity of TMD and 2D contrast studies were 100% and 75%, respectively, with specificity of 96% and 100%. Greater than 10 bubbles on a single beat of the resting contrast TMD identified patients with a maximum resting TEE PFO opening diameter >2 mm with 78% sensitivity and 100% specificity. There was a strong correlation (r2 = 0.72, p<0.01) between the TMD acoustic power and PFO opening diameter. CONCLUSIONS: Transmitral Doppler is a sensitive and specific method for TTE PFO detection that allows quantification of right to left bubble passage and may obviate the need for TEE in many patients after stroke.

Adult↗
PubMed1999

An individualized, sensitive frequency range for early detection of ototoxicity.

OBJECTIVE: The aim of this study was to identify auditory frequencies at which serial threshold testing would provide the greatest sensitivity for early detection of ototoxicity. The overall objective is to develop a more time-efficient ototoxicity monitoring protocol. DESIGN: Threshold data were analyzed from 370 hospitalized patients treated with aminoglycoside antibiotics (AMGs) or cisplatin (CDDP) who received serial auditory monitoring before, during, and after treatment at conventional (0.25 to 8 kHz) and high (9 to 20 kHz) frequencies. RESULTS: For patients showing hearing changes due to ototoxicity, a frequency range was identified for its apparent high sensitivity to initial ototoxicity. This sensitive range is identified according to an individual's hearing threshold configuration, and is, therefore, unique for each patient. The range consists of five frequencies, generally separated by 1/6 octave, e.g., 8, 9, 10, 11.2, and 12.5 kHz. To determine frequencies and combinations of frequencies that were most often involved in ototoxicity detection, threshold data in the sensitive range were analyzed in detail. This analysis suggests that patients receiving treatment with AMG or CDDP can be monitored for hearing thresholds at only five frequencies, resulting in an 84% detection rate for AMG and 94% for CDDP compared with monitoring at all conventional and high frequencies. CONCLUSIONS: This comprehensive analysis supports earlier observations that a sensitive, limited frequency range exists in which serial threshold monitoring will provide early warning of ototoxicity before effects in the speech frequency range. This finding is now being evaluated in a prospective investigation.

Aminoglycosides↗
PubMed2004

Object motion: a world view.

Moving objects are detected by virtue of their shifting image on the retina. But to know how objects are moving in the world, we must take into account the rotation of our eyes, as well as the rotation of our head. A recent paper describes neurons that carry out this computation.

Eye Movements↗
PubMed2004

Detection of pathogens from periodontal lesions.

OBJECTIVE: To comparatively detect A. actinomycetemcomitans and F. nucleatum from periodontal and healthy sites. METHODS: Subgingival clinical samples from 50 periodontitis adult patients and 50 healthy subjects were analyzed. Both organisms were isolated using a trypticase soy agar-bacitracin-vancomycin (TSBV) medium and detected by PCR. Conventional biochemical tests were used for bacteria identification. RESULTS: A. actinomycetemcomitans and F. nucleatum were isolated in 18% and 20% of the patients, respectively, and in 2% and 24% of healthy subjects. Among A. actinomycetemcomitans isolates, biotype II was the most prevalent. Primer pair AA was 100% sensitive in the detection of A. actinomycetemcomitans from both subject groups. Primers ASH and FU were also 100% sensitive to detect this organism in healthy subject samples. Primer pair FN5047 was more sensitive to detect F. nucleatum in patients or in healthy samples than primer 5059S. Primers ASH and 5059S were more specific in the detection of A. actinomycetemcomitans and F. nucleatum, respectively, in patients and in healthy subject samples. CONCLUSIONS: PCR is an effective tool for detecting periodontal pathogens in subgingival samples, providing a faster and safer diagnostic tool of periodontal diseases. The method's sensitivity and specificity is conditioned by the choice of the set of primers used.

Adult↗
PubMed2006

Detection of novel papillomaviruslike sequences in paraffin-embedded specimens of invasive and in situ squamous cell carcinomas from cats.

OBJECTIVE: To detect and partially characterize papillomavirus (PV) DNA in squamous cell carcinoma (SCC) tumor specimens from cats. SAMPLE POPULATION: 54 formalin-fixed paraffinembedded skin biopsy specimens were examined. Specimens originated from Bowenoid in situ SCC (BISC; n = 21), invasive SCC (22), and skin affected by miscellaneous nonneoplastic conditions (11). PROCEDURES: Samples from each tissue block underwent DNA extraction after deparaffinization, and PCR assays were performed. Two sets of primers derived from PV E1 were used. The first set of primers was designed for the narrow-range PCR assay and was able to generate amplification products of feline PV (FePV), canine oral PV, or closely related PVs. The second set of primers was selected for the broad-range PCR assay because of its ability to amplify DNA from 64 human PVs. Sequence analysis of each amplified DNA was performed. RESULTS: 1 of the 21 specimens of BISC was positive for PV DNA on the basis of narrow-range PCR assay results, whereas all the other specimens (BISC, invasive SCC, and controls) had negative results for PV DNA. In contrast, 5 of 21 BISC specimens and 4 of 22 invasive SCC specimens were positive for PV DNA on the basis of broad-range PCR assay results. Sequence analysis revealed that only 1 specimen was infected by a virus closely related to classic FePV. In the 8 other specimens positive for PV DNA, DNA of unknown PVs was uncovered. CONCLUSIONS AND CLINICAL RELEVANCE: Bowenoid in situ SCC and invasive SCC of cats may be associated with PVs of genetic diversity.

Animals↗
PubMed2001

[Pathogen detection in the myocardial lesion using in situ RT-PCR in mice induced by experimental CoxB(3m) virus infection].

OBJECTIVE: To detect the location of pathogens in myocardium using in situ RT-PCR technique in order to study the pathogenetic course of the myocardial lesion induced by CoxB(3m) virus infection in mice. METHODS: (1) Thirty and fifty Balb/c mice were used respectively to establish the acute and chronic CoxB(3m) infected models, with another 25 healthy mice as the controls; (2) KS400 image analysis system (Germany) was used to measure the cardiac chamber area and the left ventricular wall thickness of the chronic infected mice and the controls; (3) CoxB(3m) virus in myocardial tissue was detected using in situ RT-PCR by direct incorporated technique which employed nucleotide labeling by anti-digoxin antibody and bonded with alkaline phosphatase (anti-dig-AKP method). RESULTS: Picture analysis indicated that the left ventricular chamber area was enlarged and the left ventricular wall was thinner in the chronic repeated virus infected models than those of the controls. With in situ RT-PCR, positive signals for Coxsackie virus B(3m) RNA were detected not only in the myocardium of the acute Balb/c mice models but also in the myocardium of the chronic mice models. CONCLUSION: Coxsackie virus B(3m) is able to induce pathologic lesions by exhibiting positive CVB-RNA signals in both acute and chronic models in mice. In the chronic experimental models, the cardiac chamber is enlarged while the ventricular wall is thinned which demonstrates the association with persistent infection of Coxsackie virus B(3m) virus.

Animals↗
PubMed2003

[Detection of COL1A1/PDGFB fusion transcripts in dermatofibroscoma protuberans by revers transcriptase-polymerase chain reaction using paraffin-embedded tissues].

OBJECTIVE: To detect the COL1A1/PDGFB fusion transcripts and discuss its clinicopathological significance in dermatofibroscoma protuberans. METHODS: Formalin fixed, paraffin-embedded tumor specimens from 12 patients with DFSP were reviewed by light microscope and the expression of COL1A1/PDGFB mRNA resulting from the reciprocal translocation t(17;22) (q22;q13.1) was detected by one-step revers transcriptase-polymerase chain reaction. The following tumor specimens were included as controls: 2 fibrosarcoma, 2 malignant fibrous histocytoma, 3 leiomyosarcoma, 1 dermarofibroma and 1 nerve shealth tumor. RESULTS: The COL1A1/PDGFB fusion transcripts were detected in 8 (67%) of 12 samples from patients with DFSP. Nucleotide sequence analysis using the PCR products confirmed that different regions of the COL1A1 gene, respectively, were fused with of PDGFB gene. No COL1A1/PDGFB fusion transcripts were detected in the control tumors. CONCLUSION: Detection of specific COL1A1/PDGFB fusion transcripts in DFSP will help to diagnose the nature of DFSP and research the mechanism of its molecular histogenesis.

Adolescent↗
PubMed2004

[Quantitative detection of SARS-CoV RNA in excreta and oropharyngeal washing fluid from convalescence patients with severe acute respiratory syndrome].

OBJECTIVE: To detect the severe acute respiratory syndrome coronavirus (SARS-CoV) RNA in excreta and oropharyngeal washing fluid (OWF) from the convalescence SARS patients, and to determine whether convalescence patients carry the SARS-CoV, and whether having infectivity. METHODS: Totally 531 samples (including urine, stool, and OWF) were collected from 177 convalescence patients with positive SARS antibody which were confirmed by ELISA method. Real-time quantitative PCR was performed to detect the RNA of the SARS-CoV, and results were analyzed by SPSS analysis software. RESULTS: There were 49 (27.7%) cases of SARS-CoV RNA detection positive in 177 patients, including 31 (17.5%) cases with one sample positive, 14 (7.9%) cases with two samples positive, and 4 (2.3%) cases with three samples positive. The positive rates of urine, stool, and OWF were 14.7% (26/177), 11.9% (21/177), and 13.6% (24/177), respectively. The quantity of SARS-CoV RNA in samples was 100-47 000 copies/ml. No significant difference was found among urine, stool, and OWF on the difference grade quantity of SARS-CoV RNA quantity. CONCLUSIONS: About 10% convalescence SARS patients might still carry the SARS-CoV in hospitalization. The detection of SARS-CoV RNA by real-time quantitative PCR may become a laboratory examination warranty for convalescence SARS patients to discharge hospital and relieve separation.

Adolescent↗
PubMed2001

Concurrent processing of spatial relations and objects in two cerebral hemispheres.

This study examined hemispheric asymmetry for concurrent processing of object and spatial information. Participants viewed two successive stimuli, each of which consisted of two digits and two pictures that were randomly located and judged them as identical or different. A sample stimulus was presented in a central visual field, followed by a matching stimulus presented briefly in a left or right visual field. The matching stimuli were different from the sample stimuli with respect to the object (digit or picture) or spatial (locations or distances of items) aspect. No visual field asymmetry was found in the detection of object change. However, a left visual field advantage was found in the detection of spatial change. This result can be explained by the double filtering by frequency theory of Ivry and Robertson, who asserted that the left hemisphere has a bias for processing information contained in relatively high spatial frequencies whereas the right hemisphere has a bias for processing information contained in relatively low spatial frequencies. Based upon this evidence, the importance of interhemispheric integration for visual scene perception is discussed.

Adult↗
PubMed2001

Variability in the levels of PML-RAR alpha fusion transcripts detected by the laboratories participating in an external quality control program using several reverse transcription polymerase chain reaction protocols.

BACKGROUND AND OBJECTIVES: The detection of PML-RAR by reverse transcription (RT) polymerase chain reaction (PCR) in acute promyelocytic leukemia (APL) patients who are in hematologic remission influences therapeutic decision making in several trials. In the light of this, the Spanish group has recently designed an external quality assessment program (EQAP) of RT-PCR detection of PML-RAR, which includes a study of sensitivity of the participating laboratories. DESIGN AND METHODS: Eighteen laboratories were involved in the program. Ten laboratories followed the method of Biondi et al., 5 employed that of Borrow et al. and the 3 remaining used other protocols. The sensitivity was studied in five rounds of quality control. The first two shipments consisted of dilutions of NB4 RNA into non-APL RNA. The third round consisted of serial dilutions of the NB4 cell line into HL60 cells. The fourth and five rounds consisted of plasmid dilutions containing the bcr1 and bcr3 PML-RAR isoforms. RESULTS: The results showed that the distinct methods allow detection of the PML-RAR hybrid up to a dilution of 10(-4), and exceptionally, up to 10(-5). The laboratories following the method of Biondi et al. usually detected the 10(-3) dilution and less frequently the 10(-4) one, whereas those using other methods usually detected PML-RAR transcript in the 10(-4) dilution, and less commonly in the 10(-5) dilution. However, each of the PCR methods used by EQAP participating laboratories successfully detected at least 50 copies of PML-RAR alpha fusion transcript in plasmid dilution controls. INTERPRETATION AND CONCLUSIONS: The results point to heterogeneous sensitivity amongst participating laboratories. This may reflect differences in methodology, although variations in sample quality may also account for discrepant findings.

Humans↗