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PubMed1997

[The effect of periodontal initial therapy on detection frequency of Bacteroides forsythus, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans--detection by DNA probe and PCR methods].

The purpose of this study was to examine microbiological changes after periodontal initial therapy by DNA probe and polymerase chain reaction (PCR) assays in addition to clinical evaluation. Twenty-six periodontitis patients, 10 males and 16 females, were selected. All subjects were treated by scaling and root planing following oral hygiene instruction. Bacterial samples were collected with paper points from a total of 104 sites, 4 sites in each patient, at baseline and after initial therapy. DNA probe analysis was used to monitor Bacteroides forsythus and Porphyromonas gingivalis. PCR analysis was performed to identify the presence of Actinobacillus actinomycetemcomitans. The initial therapy resulted in significant clinical improvements as assessed by clinical parameters including pocket depth, attachment level and bleeding on probing. After the therapy, the detection frequency of B. forsythus and P. gingivalis was significantly reduced, but the detection frequency of A. actinomycetemcomitans did not significantly decreased. These results indicated that initial therapy can bring about elimination of B. forsythus and P. gingivalis by means of conventional techniques, but not of A. actinomycetemcomitans. When complete elimination of these microorganisms was obtained, significant clinical improvement was found. Monitoring the changes of three bacteria may provide a more effective modality.

Adolescent↗

PubMed2005

Fetal magnetic resonance imaging enhances detection of spinal cord anomalies in patients with sonographically detected bony anomalies of the spine.

OBJECTIVE: Although fetal magnetic resonance imaging (MRI) is being increasingly used to evaluate sonographically suspected abnormalities, its utility in the evaluation of the spinal canal is not well studied. Because it is not susceptible to the limitations of fetal position, oligohydramnios, and shadowing from bony structures, we hypothesize that fetal MRI is better suited to assess the contents of the spinal canal compared with prenatal sonography. The purpose of this investigation was to determine whether fetal MRI could detect spinal abnormalities in cases in which they had not been originally suspected on prenatal sonography. METHODS: Fetal spine MR images were retrospectively reviewed over a 42-month period. Corresponding sonographic images were then rereviewed to determine whether there were findings in retrospect that might have suggested the cord abnormalities. Cases of myelomeningocele were counted as a spinal cord abnormality only if fetal MRI showed a cord anomaly other than the myelomeningocele. RESULTS: Of 33 cases referred for bony anomalies of the spine, fetal MRI showed additional abnormalities involving the spinal cord in 3 patients. These included diastematomyelia in 2 cases and segmental spinal dysgenesis in the third case. One case of diastematomyelia occurred in association with a lumbosacral myelomeningocele. The spinal cord anomalies were not visible on any of the prenatal sonograms, even in retrospect. CONCLUSIONS: Additional spinal cord anomalies were detected in 10% of cases reviewed. Fetal MRI can be useful in assessing the spinal cord in fetuses with bony spinal anomalies. Our findings suggest that fetuses with sonographically diagnosed bony abnormalities of the spine may benefit from further evaluation with fetal MRI.

Female↗

PubMed2000

Immunohistochemical detection of CD10 in paraffin sections of hematopoietic neoplasms: a comparison with flow cytometry detection in 56 cases.

Paraffin-section immunohistochemistry with heat-induced epitope retrieval using a newly characterized monoclonal antibody (clone 56C6) against the CD10 antigen was performed on 56 hematopoietic tumors previously studied for CD10 expression by flow cytometry. The cases included 33 precursor B-lymphoblastic leukemias, 10 acute myeloid leukemias, five precursor T-lymphoblastic leukemias, five follicular lymphomas, and three Burkitt cell leukemias. Forty of the 56 cases were CD10 positive by flow cytometry studies, including all five follicular lymphomas (100%); 30 of 33 (91%) cases of precursor B-lymphoblastic leukemias, two of three (66%) cases of Burkitt cell leukemias, two of five (40%) cases of precursor T-lymphoblastic leukemias, and none of the 10 cases of acute myeloid leukemia. Thirty-nine of the 40 (97%) flow cytometric CD10-positive cases also expressed CD10 by immunohistochemistry in formalin- or B5-fixed, paraffin-embedded tissue, with only one case of precursor B-lymphoblastic leukemia being positive by flow cytometry and negative by immunohistochemistry. The 16 CD10-negative flow cytometry specimens were all also negative by immunohistochemistry. Thirty-seven CD10 immunohistochemistry positive cases showed a diffuse membranous staining pattern and two cases demonstrated a Golgi staining pattern. The fixation methods (10% neutral buffered formalin versus B5) and decalcification did not affect the CD10 immunostaining results. This study demonstrates that the new CD10 monoclonal antibody clone 56C6 is a reliable marker for detection of CD10 antigen expression in formalin-and B5-fixed paraffin-embedded tissue after heat-induced epitope retrieval when compared with flow cytometry detection of fresh tissue samples.

Antibodies, Monoclonal↗

PubMed2001

Enzymatic mutation detection method evaluated for detection of p53 mutations in cDNA from breast cancers.

BACKGROUND: Rapid, reproducible, and easily run methods with high sensitivity and specificity are required for mutation screening of clinical samples. We evaluated the Enzymatic Mutation Detection (EMD(TM)) method by analysis of archival cDNA from 203 breast cancer patients and comparison with results of cDNA-based sequencing of the tumor suppressor gene p53. METHODS: The EMD technology uses the T4 endonuclease VII, which cleaves double-stranded DNA at sites where a DNA mismatch is present because of mispairing or an insertion/deletion of nucleotides. The EMD analyses were carried out by dividing the p53 gene into two overlapping fragments that were analyzed separately. After PCR amplification, the fragments were hybridized with wild-type p53 and subsequently exposed to the EMD enzyme. Cleavage products were analyzed and scored using an ALF(TM) automated DNA sequencer and ALFwin Fragment Analyzer software (VER: 1.02). RESULTS: The EMD technique had sensitivities of 45% and 64% and specificities of 83% and 84% for the two fragments, respectively. Patients with EMD-positive, wild-type p53 tumors had a survival similar to that of patients with EMD-negative, wild-type p53 tumors. Node-positive patients with p53 mutated tumors according to sequencing had a statistically significantly worse overall survival than those with p53 wild-type tumors (P = 0.016), whereas this difference in survival was not detected when p53 status was determined with EMD (P = 0.47). CONCLUSIONS: EMD had insufficient sensitivity for consideration in screening for the p53 gene in this archival material. Sequencing must still be considered as the standard procedure.

Breast Neoplasms↗

PubMed2000

Comparison of different sperm detection methods: amplification of fragment 3 of amelogenin gene (AMY 3), acid phosphatase detection, microcristalline choline assay and microscopic examination.

Different methods of male material detection in vaginal swabs after the intercourse are compared and evaluated. Advantages of PCR technique in sperm detection are presented and discussed.

Acid Phosphatase↗

PubMed2002

Telephotoscreening to detect retinopathy of prematurity: preliminary study of the optimum time to employ digital fundus camera imaging to detect ROP.

PURPOSE: Labor-intensive screening of infants in the neonatal intensive care units is the only way presently to detect retinopathy of prematurity (ROP). Our purpose is to determine if RetCam 120 images (Massie Research Laboratories, Inc, Dublin, Calif), acquired by a neonatal nurse, can be used to screen for ROP by performing 2 screening sessions, at 32 to 34 weeks' (examination 1) and 38 to 40 weeks' (examination 2) postconceptional age. METHODS: RetCam examinations were performed by a nurse on infants at examination 1 and examination 2 intervals. At the same time, an examination was performed by an experienced ophthalmologist. Masked readers evaluated the digital images for the presence of ROP and, if ROP was present, estimated the risk of that eye progressing to prethreshold or threshold disease. The data were compared to the eye's clinical course. RESULTS: A total of 46 eyes were assessed at examination 1 and 50 eyes at examination 2 from July 1, 1999, to December 15, 1999. For detecting ROP, the sensitivity and specificity were 46% and 100% for examination 1 and 76% and 100% for examination 2. Sensitivity and specificity of predicting prethreshold was 64% and 97%, respectively, for examination 1 and 2. Sensitivity for predicting ROP threshold at examination 1 was 0% (only 1 photo was available for grading of sensitivity) and specificity for predicting ROP threshold at examination 1 was 95%. At examination 2, sensitivity and specificity were 100%. CONCLUSION: The RetCam examination had insufficient sensitivity to be recommended as a substitute for indirect ophthalmoscopy in screening for ROP. Reasons for low sensitivity are the technical limitation of the camera design itself, which creates difficulty in photographing the peripheral retina in small eyes, and the need for a lid speculum better adapted to the contact camera optical system design. Both of these issues are being addressed as part of an ongoing project to study the feasibility of employing telemetry of digital fundus images from remote, underserved neonatal intensive care units to ophthalmologists capable of diagnosing ROP.

Diagnostic Techniques, Ophthalmological↗

PubMed2002

[Management of mucocutaneous herpes simplex virus infections in immunocompetent patients: signification and limits of antigen detection culture methods and antibody detection].

Herpes simplex virus (HSV) infections are very common and may present various manifestations. Mostly asymptomatic, often mild, these infections may become life-threatening, specially in neonates. The acute and rapid diagnosis of HSV infections can prevent infections in these patients and is also helpful to confirm clinical diagnosis. The sensitivity of "classical" diagnosis methods, such as culture and antigen detection by immunofluorescence and ELISA, is highly dependent on the quality of the sample. Antigen detection techniques give results in short delays, compatible with the initiation of antiviral treatment; however their sensitivity decreases when lesions get older and they are not convenient for the diagnosis of asymptomatic infections. Isolation of HSV on cell culture remains the gold standard, because it is easy to perform and exhibits good sensitivity whatever the stage of the lesions; the isolation of the virus is required for HSV typing and to determine susceptibility to antiviral agents in case of clinical resistance to the treatment. Serodiagnosis is helpful to determine the immune status of a patient and to distinguish between primary infection and recurrence. It is now possible to differentiate HSV1 and HSV2 antibodies; the benefit of type-specific serology is clear for epidemiological surveys but may be discussed for individual follow-up, specially for genital herpes, both for the diagnosis and the prevention of HSV infections.

Enzyme-Linked Immunosorbent Assay↗

PubMed2003

[Application of PCR to Haemophilus influenzae typing. I. PCR standardization for bexA gene fragment detection and for detection of DNA fragment specific for H.influenzae type B].

PCR standardization was performed in order to detect a fragment of bexA gene, which is presented in all capsulate H. influenzae isolates, and a DNA fragment specific for H. influenzae type b. Implementing of such PCR into H. influenzae typing may be very useful in cases of isolates from clinical material for which serotyping alone gives unclear results. Standardization of PCR detecting DNA fragments specific for all capsular types will enable to perform complete typing of H. influenzae isolates.

Bacterial Typing Techniques↗

PubMed2004

Prostate-specific antigen and free prostate-specific antigen in the early detection of prostate cancer: do combination tests improve detection?

BACKGROUND: The combined use of free and total prostate-specific antigen (PSA) in early detection of prostate cancer has been controversial. This article systematically evaluates the discriminating capacity of a large number of combination tests. METHODS: Free and total PSA were analyzed in stored serum samples taken prior to diagnosis in 429 cases and 1,640 controls from the Physicians' Health Study. We used a classification algorithm called logic regression to search for clinically useful tests combining total and percent free PSA and receiver operating characteristic analysis and compared these tests with those based on total and complexed PSA. Data were divided into training and test subsets. For robustness, we considered 35 test-train splits of the original data and computed receiver operating characteristic curves for each test data set. RESULTS: The average area under the receiver operating characteristic curve across test data sets was 0.74 for total PSA and 0.76 for the combination tests. Combination tests with higher sensitivity and specificity than PSA > 4.0 ng/mL were identified 29 out of 35 times. All these tests extended the PSA reflex range to below 4.0 ng/mL. Receiver operating characteristic curve analysis indicated that the overall diagnostic performance as expressed by the area under the curve did not differ significantly for the different tests. CONCLUSIONS: Tests combining total and percent free PSA show modest overall improvements over total PSA. However, utilization of percent free PSA below a PSA threshold of 4 ng/mL could translate into a practically important reduction in unnecessary biopsies without sacrificing cancers detected.

Aged↗

PubMedDate not supplied

[Detection of single base polymorphism in p53 gene by ligase detection reaction and rolling circle amplification on microarrays].

We combined three modern technologies of single base polymorphism detection in human genome: ligase detection reaction, rolling circle amplification and IMAGE hydro-gel microarrays. Polymorphism in target DNA was tested by selective ligation on microarray. Product of the ligase reaction was determined in microarray gel pads by rolling circle amplification. Two different methods were compared. In first, selective ligation of short oligonucleotides immobilized on microarray was used with subsequent amplification on preformed circle probe ("common circle"). The circle probe was designed especially for human genome research. In second variant, allele-specific padlock probes that may be circularized by selective ligation were immobilized on microarray. Polymorphism of codon 72 in human p53 gene was used as a biological model. It was shown that LDR/RCA on microarray is a quantitative reaction and gives high discrimination of alleles. Principles and perspectives of selective ligation and rolling circle amplification are being discussed.

DNA Ligases↗

PubMed2006

Novel cryptic chromosomal rearrangements detected in acute lymphoblastic leukemia detected by application of new multicolor fluorescent in situ hybridization approaches.

Routine cytogenetic analysis provides important information on diagnostic and prognostic relevance for hematological malignancies. However, it is often difficult to obtain good karyotypes, especially of cells from cases with acute lymphoblastic leukemia (ALL) because of poor morphology and spreading. Thus, detailed karyotyping can be hampered and even in case of a 'normal karyotype' according to banding cytogenetics doubts remain if the result is reliable. In order to address this problem a series of 37 ALL cases without any detectable numerical or structural chromosomal defects was selected and studied by two recently developed multicolor fluorescence in situ hybridization (FISH) approaches: 1) multitude multicolor banding (mMCB) is a FISH-banding technique, which allows the analyses of inter- and intra-chromosomal rearrangements of the whole human karyotype in one single experiment; 2) chromosome-specific subcentromere/subtelomere-specific multicolor (subCTM-)FISH applies locus-specific subtelomeric and subcentromic probes and enables the characterization of the subtelomeric and peri-centric regions of the chromosomes, not analyzable by other FISH-approaches. Thus, we detected the following recurrent cryptic chromosomal aberrations: del(12)(pter) [8 cases], del(9)(qter) [3 cases], and del(11)(pter) [2 cases]. Moreover, cryptic changes in additional nine subtelomeric and in two subcentromeric regions were observed one time, each. In summary, mMCB and subCTM were proven to be powerful methods in the screening for new cryptic chromosomal aberrations, which considerably increased the accuracy of cytogenetic diagnosis.

Adolescent↗

PubMed2006

[Comparison between rapid detection method of enzyme substrate technique and multiple-tube fermentation technique in water coliform bacteria detection].

OBJECTIVE: To compare between rapid detection method of enzyme substrate technique and multiple-tube fermentation technique in water coliform bacteria detection. METHODS: Using inoculated and real water samples to compare the equivalence and false positive rate between two methods. RESULTS: Results demonstrate that enzyme substrate technique shows equivalence with multiple-tube fermentation technique (P = 0.059), false positive rate between the two methods has no statistical difference. CONCLUSION: It is suggested that enzyme substrate technique can be used as a standard method for water microbiological safety evaluation.

Bacteriological Techniques↗

PubMedDate not supplied

Early detection of ovarian cancer: background, rationale, and structure of the Yale Early Detection Program.

Ovarian cancer has received national attention as a highly virulent disease. Its lack of early warning symptoms and the failure to develop highly sensitive screening tests have led some physicians to recommend prophylactic oophorectomies to women with relatives who have had ovarian cancer. Others have recommended routine screening of otherwise normal women for CA 125, a circulating tumor marker, and ultrasound examinations. Each of these techniques is associated with substantial false-positive rates that could lead to unnecessary surgery. A review of epidemiologic data suggests that familial ovarian cancer kindreds are rare, but women with first-degree relatives who have had ovarian cancer have a significant risk themselves for developing ovarian cancer. In addition, women with a great number of ovulatory cycles are at an increased risk for the disease. Circulating tumor markers are frequently elevated in women with advanced ovarian cancer, but their value in early detection of ovarian cancer has yet to be established. Advances in endovaginal ultrasound and color Doppler flow technology have significantly improved our ability to assess pelvic organs. This article presents the background, rationale, and structure of the Yale Early Detection Program for ovarian cancer, whose goals are to identify the best techniques for diagnosing ovarian cancer in an early stage, to determine the frequency with which such tests should be employed, to assess false-positive results, and to identify women who might benefit from prophylactic oophorectomies.

Biomarkers, Tumor↗

PubMedDate not supplied

Early detection of ovarian cancer: preliminary results of the Yale Early Detection Program.

Eighty-four women at high risk for ovarian cancer by having first-degree relatives with epithelial ovarian cancer participated in a newly established, early ovarian cancer detection program at Yale University. Participants were to be evaluated with physical examinations and circulating tumor markers at entry and every six months thereafter. Endovaginal ultrasound and color Doppler flow studies were to be performed at three and nine months following entry into the program. In addition, women were encouraged to follow American Cancer Society guidelines for mammography. Stool was checked for occult blood. Endometrial sampling was offered to post-menopausal women. No participant has developed an ovarian cancer since entering the program. One woman has been diagnosed to have breast cancer. False-positive levels of circulating tumor markers (CA 125, 4/84 [4.8 percent]; lipid-associated sialic acid in plasma, 13/84 [15.5 percent]; NB/70K, 4/84 [4.8 percent]; and urinary gonadotropin fragment, 1/65 [1.5 percent]) were observed on entry into the program. Low resistive indices (less than 0.5) were documented in 8/91 (8.8 percent) ovaries studied by the color Doppler flow technique. One participant underwent a laparotomy based on a false-positive endovaginal ultrasound examination. Tests now being employed in community practice have a high likelihood of being associated with false-positive results. Therapeutic interventions based on isolated abnormal tumor markers or ultrasound studies obtained from women with family histories of ovarian cancer may lead to inappropriate surgery. It is necessary for cancer centers to develop expertise in ovarian cancer detection techniques to advise physicians in their geographic areas appropriately about the significance of the abnormal screening test.

Adult↗

PubMed1991

[Detection of antibiotics in biological materials for purposes of toxicological analysis. I. Methodologic study for the detection of tetracycline].

A method of the identification of tetracycline antibiotics in urine and gastric content, suitable for the routine toxicological analysis, was worked out. For the isolation, the extraction with ethyl acetate is recommended. The TLC method on Silufol layers impregnated with an aqueous solution of the disodium salt of ethylenediaminetetraacetic acid (Na2EDTA), in the concentration of 0.1 mol/l and pH value of 5 or 7.4 is used for the identification. The mobile phase chloroform--methanol--Na2EDTA 55:30:5 is suitable for the development of these layers; UV light of the wavelength 254 or 366 nm and Fast Blue B reagent are used for the detection. The conditions of the TLC (various modifications of the layer, mobile phase and detection) and the extraction solvents suitable for the isolation are discussed.

Chromatography, Thin Layer↗

PubMed1989

Immunoassay detection of drugs in racing horses. IX. Detection of detomidine in equine blood and urine by radioimmunoassay.

Detomidine is a potent non-narcotic sedative agent which is currently in the process of being approved for veterinary clinical use in the United States. Since no effective screening method in horses is available for detomidine, we have developed an 125I radioimmunoassay for detomidine in equine blood and urine as part of a panel of tests for illegal drugs in performance horses. Our 125I radioimmunoassay has an I-50 for detomidine of approximately 2 ng/ml. Our assay shows limited cross-reactivity with the pharmacodynamically similar xylazine, but does not cross-react with acepromazine, epinephrine, haloperidol or promazine. The plasma kinetic data from clinical (greater than or equal to 5 mg/horse) as well as sub-clinical doses indicate first-order elimination in a dose-dependent manner. Within the first 30 minutes after intravenous (IV) administration of 30 mg/horse, plasma levels peak at approximately 20 ng/ml and then decline with an apparent plasma half-life of 25 minutes. Diuresis can occur with administration of clinical doses of detomidine and this effect was accounted for in the analysis of urine samples. Using this method, administration of 30 mg/horse can be readily detected in equine urine for up to 8 hours after IV injection. Additionally, doses as low as 0.5 mg/horse can be detected for short periods of time in blood and urine with use of this assay. Utilization of this assay by research scientists and forensic analysts will allow for the establishment of proper guidelines and controls regarding detomidine administration to performance horses and assurance of compliance with these guidelines.

Analgesics↗

PubMedDate not supplied

Anatomic viral detection is automated: the application of a robotic molecular pathology system for the detection of DNA viruses in anatomic pathology substrates, using immunocytochemical and nucleic acid hybridization techniques.

This paper presents the first automated system for simultaneously detecting human papilloma, herpes simplex, adenovirus, or cytomegalovirus viral antigens and gene sequences in standard formalin-fixed, paraffin-embedded tissue substrates and tissue culture. These viruses can be detected by colorimetric in situ nucleic acid hybridization, using biotinylated DNA probes, or by indirect immunoperoxidase techniques, using polyclonal or monoclonal antibodies, in a 2.0-hour assay performed at a single automated robotic workstation.

Antigens, Viral↗

PubMed1987

[Detection of an inhibitor of the causative agent of plague active against strains of the plague microbe from an homologous focus. The effect of lithium chloride on detecting the inhibitor].

A nutrient medium containing 0.7 per cent of lithium chloride was tested and Yersinia pestis inhibitor was detected in 83.3 per cent of the strains on the growth tufts of 8 indicator strains isolated from a homologous focus. Lowering the lithium chloride concentration in the medium up to 0.6 per cent promoted detection of the inhibitor in a larger number of the tested strains (93.3 per cent). But even on this medium none of the indicator strains formed overall growth tufts. The subcultures of the indicator strains resistant to lithium chloride formed overall growth tufts on agar plates with lithium chloride and were sensitive to the inhibitor of the tested strains from the central Caucasus. However, during storage they lost their sensitivity to the inhibitor which did not permit their use as the test cultures.

Antibiosis↗