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PubMed1998

Comparison of the ABI 7700 system (TaqMan) and competitive PCR for quantification of IS6110 DNA in sputum during treatment of tuberculosis.

Mycobacterium tuberculosis can persist in sputum for long periods of time after the initiation of antituberculosis chemotherapy. The purpose of this study was to determine whether quantitative estimates of M. tuberculosis DNA in sputum correlate with the numbers of viable bacilli and thus measure the therapeutic response of patients during treatment. Two methods of M. tuberculosis DNA quantification were examined by using DNA isolated from sputum specimens serially collected during the course of chemotherapy. A competitive PCR assay was compared to an automated system of real-time quantification with the ABI Prism 7700 Sequence Detection System (TaqMan). The ABI 7700 system uses standard PCR in conjunction with a fluorogenic probe in which the intensity of fluorescence is proportional to the amount of target DNA present. The results showed that both PCR systems are reproducible and accurate. The amounts of M. tuberculosis DNA quantified in sputum corresponded well with the numbers of acid-fast bacilli (AFB) counted by microscopy. Before initiation of antituberculosis therapy, measures of AFB, M. tuberculosis DNA, and cultivable bacilli were similar, suggesting that quantification of DNA is a good method for measuring the initial bacillary load. However, the rate of disappearance of both AFB and M. tuberculosis DNA did not correlate with the decline in cultivable bacilli in the specimen; therefore, these tests are not appropriate for monitoring treatment efficacy.

Antitubercular Agents↗
PubMed1999

Nucleic acid-based cross-linking assay for detection and quantification of hepatitis B virus DNA.

A nucleic acid photo-cross-linking technology was used to develop a direct assay for the quantification of hepatitis B virus (HBV) DNA levels in serum. Cross-linker-modified DNA probes complementary to the viral genomes of the major HBV subtypes were synthesized and used in an assay that could be completed in less than 6 h. The quantification range of the assay, as determined by testing serial dilutions of Eurohep HBV reference standards and cloned HBV DNA, was 5 x 10(5) to 3 x 10(9) molecules of HBV DNA/ml of serum. Within-run and between-run coefficients of variation (CVs) for the assay were 4. 3 and 4.0%, respectively. The assay was used to determine HBV DNA levels in 302 serum samples, and the results were compared to those obtained after testing the same samples with the Chiron branched-DNA (bDNA) assay for HBV DNA. Of the samples tested, 218 were positive for HBV DNA by both assays and 72 gave results below the cutoff for both assays. Of the remaining 12 samples, 10 were positive for HBV DNA by the cross-linking assay only; the 2 other samples were positive by the bDNA assay only. Twenty-eight samples had to be retested by the bDNA assay (CV, >20% between the results obtained from the testing of each sample in duplicate), whereas only three samples required retesting by the cross-linking assay. The correlation between the HBV DNA levels, as measured by the two tests, was very high (r = 0.902; P = 0.01). We conclude that the cross-linking assay is a sensitive and reproducible method for the detection and quantification of HBV DNA levels in serum.

Cross-Linking Reagents↗
PubMed1999

Rapid and sensitive quantification of Borrelia burgdorferi-infected mouse tissues by continuous fluorescent monitoring of PCR.

The quantity of Borrelia burgdorferi organisms in tissue samples is an important determinant for infection studies in the mouse model of Lyme disease. This report presents the development of a rapid and sensitive external-standard-based PCR assay for the absolute quantification of B. burgdorferi in mouse tissue samples. The assay uses a double-stranded DNA dye to continuously monitor product formation and in less than an hour was able to quantify samples ranging up to 6 log units in concentration. The PCR efficiencies of the sample and the standard were matched by using a standard composed of purified B. burgdorferi chromosome mixed with tissue-matched mouse genome lacking bacterial DNA. Normalization of B. burgdorferi quantities to the mouse nidogen gene allowed comparison of B. burgdorferi numbers in samples isolated from different tissues and strains. PCR analysis of the chromosomal gene recA in cultured B. burgdorferi was consistent with a single recA per bacterium. The parameters defined in this assay should be applicable to quantification of other organisms, even infectious agents for which no ready source of DNA standard is available. In summary, this report presents a rapid external-standard-based PCR method for the quantification of B. burgdorferi in mouse DNA samples.

Animals↗
PubMed2001

Direct quantification of human cytomegalovirus immediate-early and late mRNA levels in blood of lung transplant recipients by competitive nucleic acid sequence-based amplification.

The dynamics of active human cytomegalovirus (HCMV) infection was monitored by competitive nucleic acid sequence-based amplification (NASBA) assays for quantification of IE1 (UL123) and pp67 (UL65) mRNA expression levels in the blood of patients after lung transplantation. RNA was isolated from 339 samples of 13 lung transplant recipients and analyzed by the quantitative IE1 and pp67 NASBA in parallel with pp65 antigenemia and serology. Rapid increases in IE1 RNA exceeding 10(4) copies per 100 microl of blood were associated with active infection, whereas lower levels were suggestive for abortive, subclinical viral activity. Any positive value for pp67 RNA was indicative for active infection, and quantification of pp67 mRNA did not give additional diagnostic information. The onset of IE1-positive NASBA preceded pp67 NASBA and was earlier than the pp65 antigenemia assay, confirming previous studies with qualitative NASBA. Effective antiviral treatment was reflected by a rapid disappearance of pp67 mRNA, whereas IE1 mRNA remained detectable for longer periods. Quantification of IE1 might be relevant to monitor progression of HCMV infection but should be validated in prospective studies.

Antibodies, Viral↗
PubMed2005

Detection and quantification of human immunodeficiency virus type 1 p24 antigen in dried whole blood and plasma on filter paper stored under various conditions.

The quantification of human immunodeficiency virus type 1 (HIV-1) by an assay measuring heat-dissociated (HD) p24 antigen (Ag) in specimens of whole blood and plasma stored on filter paper, and of plasma stored in tubes, was compared to HIV-1 RNA plasma levels determined by real-time reverse transcription (RT)-PCR. The stability of p24 Ag on filter paper under conditions simulating specimen transport was also evaluated. The HD p24 Ag in both plasma and whole-blood specimens stored on filter paper correlated with plasma HIV-1 RNA levels (Spearman rank rho = 0.74 [P < 0.0001] and rho = 0.56 [P = 0.0001], respectively). The sensitivity of the HD p24 Ag assay was similar when plasma and whole blood on filter paper were contrasted to the real-time RT-PCR assay (80% versus 82.5% and 78.6% versus 83.3%, respectively). However, while the specificity of the HD p24 Ag assay of plasma on filter paper was 100%, the specificity was diminished in whole-blood specimens. The storage of specimens on filter paper for 2 weeks at 37 degrees C, 24 degrees C, or 0 degrees C did not alter the detection or quantification of HD p24 Ag. These results suggest that transport and storage of plasma on filter paper and quantification of HD p24 Ag may be a reliable method for HIV-1 load monitoring.

Filtration↗
PubMed2003

The "X-Ray RheumaCoach" software: a novel tool for enhancing the efficacy and accelerating radiological quantification in rheumatoid arthritis.

BACKGROUND: Precise diagnosis and follow up treatment of rheumatoid arthritis (RA) requires objective quantification, which is still lacking. For this purpose, radiological analyses are considered to be the most appropriate method. OBJECTIVE: To develop computer assisted quantification software that is particularly applicable to joint scoring in rheumatic disorders. METHODS: 3914 radiographs from hands and feet of 190 patients with RA were collected, expertly examined, analysed, and statistically evaluated. Radiographs were quantified using the conventional Larsen score and the "X-Ray RheumaCoach" (XRRC) software. The XRRC is a Java stand alone application which can support and accelerate, but not fully automate, the scoring procedure in RA. The scorer can apply both the Larsen and the Ratingen-Rau scores. RESULTS: Compared with conventional scoring procedures, the XRRC software accelerated quantification time by approximately 25%. The program, which is now available on the internet free of charge, ran stably and proved to be a consistently valuable tool. CONCLUSIONS: Compared with conventional scoring methods, the XRRC software offers several advantages: (a) structured data analysis and input that minimises variance by standardisation; (b) faster and more precise calculation of sum scores and indices; (c) permanent data storing and fast access to the software's database; (d) the possibility of cross calculation to other scores; (e) "user friendly" technology and a dedicated help program; (f) fast access and data transfer through the internet if desired; and (g) reliable documentation of results in a specially designed printout.

Arthritis, Rheumatoid↗
PubMed2002

Coronary artery calcium: absolute quantification in nonenhanced and contrast-enhanced multi-detector row CT studies.

PURPOSE: (a) To determine the accuracy of multi-detector row computed tomography (CT) in the measurement of the calcium concentration in a cardiac CT calibration phantom and (b) to assess the correlation of a traditional 3-mm section width CT coronary screening protocol and a 1.25-mm section width CT angiography imaging protocol in the quantification of the absolute mass of coronary calcium in patients who underwent both coronary screening and CT angiography with a multi-detector row CT scanner. MATERIALS AND METHODS: A heart phantom containing calcified cylinders was scanned to determine calibration factors and absolute calcium mass. In 50 patients, the variability (value 1 - value 2/mean value 1 - value 2), limit of agreement (+/-2SD value 1 - value 2), and systematic error (mean value 1 - value 2) of the total amount of coronary calcium calculated at traditional 3-mm section width CT and at 1.25-mm section width CT angiography were determined. RESULTS: The correlation coefficient between the 3-mm section width, nonenhanced protocol and the 1.25-mm section width CT angiography protocol was very high (r = 0.977) and the mean variability was low (19.7%) for the absolute mass. There was a systematic error of -6.7 mg and a limit of agreement between 45.0 mg and -58.5 mg. CONCLUSION: Use of the mass quantification algorithm in combination with a calibration phantom allows accurate quantification of coronary calcium. Measurements of calcium mass obtained at 1.25-mm section width CT angiography have the best agreement with those obtained at the traditional 3-mm section width imaging protocol.

Algorithms↗
PubMed2003

Coronary artery calcium: accuracy and reproducibility of measurements with multi-detector row CT--assessment of effects of different thresholds and quantification methods.

PURPOSE: To evaluate the effects of different thresholds and quantification methods on the accuracy and reproducibility of coronary calcium measurements with multi-detector row computed tomography (CT). MATERIALS AND METHODS: A cardiac CT phantom containing predetermined calcified cylinders was scanned. Calcium volume and mass were measured at various threshold values ranging from 80 to 230 HU. In 32 patients, two consecutive CT scans were obtained, and the coronary artery calcium score, volume, and mass were measured by one observer at 130- and 90-HU thresholds. Correlation analysis and analysis of variance were performed to evaluate the measurement errors in the phantom study and the interscan variability in the clinical study. RESULTS: In the phantom, mass measurement error varied with threshold and calcium density (P <.01). Mass error was strongly correlated with volume error (r = 0.91, P <.01) but with a much smaller range. In the clinical study, interscan variability of mass measurements was significantly lower than that with other measurement methods for both patients and individual vessels. For the patients, the mean interscan variability of calcium score, volume, and mass at the 130-HU threshold was 20.4%, 13.9%, and 9.3%, respectively. For all methods, interscan variability was not significantly different between the 130- and 90-HU thresholds (P >.05). CONCLUSION: The mass measurement is more accurate, less variable, and more reproducible in coronary calcium quantification than are measurements with other algorithms. Accurate quantification of calcium in each calcified plaque may require that the threshold be set individually, depending on the calcium density.

Algorithms↗
PubMed2003

MR quantification of hepatic iron concentration.

PURPOSE: To evaluate the accuracy of magnetic resonance (MR) imaging in the quantification of hepatic iron concentration. MATERIALS AND METHODS: Between April 1999 and June 2001, 112 patients were recruited prospectively. All had undergone liver biopsy and hepatic iron concentration quantification with spectrophotometry, followed by MR imaging. MR imaging involved use of four gradient-echo sequences and one spin-echo sequence. Signal intensity (SI) was measured on images obtained with each sequence by means of regions of interest placed in the liver and paraspinal muscle to obtain the liver-to-muscle SI ratio. The relationship between hepatic iron concentration and SI ratio for each sequence was analyzed with multiple linear regression. Receiver operating characteristic analysis was performed to find the diagnostic thresholds. RESULTS: Sixty-eight patients had normal hepatic iron levels (<36 micromol/g), 23 had hemosiderosis (36-80 micromol/g), and 21 had hemochromatosis (>80 micromol/g). With all sequences, an inverse linear relationship between iron concentration and SI ratio was apparent. The authors generated a mathematic model to estimate the iron concentrations from MR imaging data (r = 0.937). For estimated concentrations of more than 85 micromol/g, the positive predictive value for hemochromatosis was 100%; for those less than 40 micromol/g, the negative predictive value for hemochromatosis was 100%. For estimated concentrations of more than 58 micromol/g, the positive predictive value for iron overload was 100%; for those less than 20 micromol/g, the negative predictive value for iron overload was 100%. CONCLUSION: MR imaging is a useful and noninvasive diagnostic tool for quantification of hepatic iron concentration.

Adult↗
PubMed2005

Intramyocellular lipid quantification: repeatability with 1H MR spectroscopy.

PURPOSE: To prospectively determine the repeatability and variability of tibialis anterior intramyocellular lipid (IMCL) quantifications performed by using 1.5-T hydrogen 1 (1H) magnetic resonance (MR) spectroscopy in healthy subjects. MATERIALS AND METHODS: Institutional review board approval and written informed consent were obtained for this Health Insurance Portability and Accountability Act-compliant study. The authors examined the anterior tibial muscles of 27 healthy subjects aged 19-48 years (12 men, 15 women; mean age, 25 years) by using single-voxel short-echo-time point-resolved 1H MR spectroscopy. During a first visit, the subjects underwent 1H MR spectroscopy before and after being repositioned in the magnet bore, with voxels carefully placed on the basis of osseous landmarks. Measurements were repeated after a mean interval of 12 days. All spectra were fitted by using Java-based MR user interface (jMRUI) and LCModel software, and lipid peaks were scaled to the unsuppressed water peak (at 4.7 ppm) and the total creatine peak (at approximately 3.0 ppm). A one-way random-effects variance components model was used to determine intraday and intervisit coefficients of variation (CVs). A power analysis was performed to determine the detectable percentage change in lipid measurements for two subject sample sizes. RESULTS: Measurements of the IMCL methylene protons peak at a resonance of 1.3 ppm scaled to the unsuppressed water peak (IMCL(W)) that were obtained by using jMRUI software yielded the lowest CVs overall (intraday and intervisit CVs, 13.4% and 14.4%, respectively). The random-effects variance components model revealed that nonbiologic factors (equipment and repositioning) accounted for 50% of the total variability in IMCL quantifications. Power analysis for a sample size of 20 subjects revealed that changes in IMCL(W) of greater than 15% could be confidently detected between 1H MR spectroscopic measurements obtained on different days. CONCLUSION: 1H MR spectroscopy is feasible for repeatable quantification of IMCL concentrations in longitudinal studies of muscle metabolism.

Adult↗
PubMed2005

Functional CT quantification of tumor perfusion after transhepatic arterial embolization in a rat model.

PURPOSE: To quantify tumor perfusion after transcatheter arterial embolization (TAE) with functional computed tomography (CT) and to validate the reproducibility of quantification measurements. MATERIALS AND METHODS: This study was conducted in accordance with an institutional animal care and use committee-approved protocol. Sixteen rats with liver tumors underwent TAE with 1 mg (group 1) or 3 mg (group 2) of polyvinyl alcohol particles. In each group, four rats underwent functional CT immediately after TAE (day 0) and four others underwent functional CT 2 days after TAE (day 2). Another four rats served as control rats. Blood flow (BF), blood volume (BV), mean transit time (MTT), and permeability-surface area product were measured by using a functional CT software program. For evaluation of reproducibility, six additional rats with mammary tumors underwent functional CT twice, with examinations 2 hours apart. The mixed-effect model was used to assess the TAE treatment effect, and the Pearson correlation test was used to determine measurement reproducibility. RESULTS: With the exception of BF in group 1 on day 2 (P = .41), BF and BV values in both groups on both days were significantly lower than BF and BV values in the control rats (with P values ranging from .018 to <.001). BF was significantly lower in group 2 than in group 1 on days 0 and 2 (P = .043 and P = .02, respectively). BV was significantly lower on day 2 than on day 0 in group 2 (P = .016). MTT was generally inversely related to BF. MTTs in group 2 on days 0 and 2 were significantly longer than those in the control rats (P < .001 and P = .03, respectively), and MTT was shorter on day 2 than on day 0 in group 2 (P = .02). Permeability-surface area product changes were similar to BF changes. There were no significant differences (P values ranged from .2 to .5) between perfusion parameters in the reproducibility study. CONCLUSION: The results of this study validate the use of functional CT in the quantification of tumor perfusion after TAE and the reproducibility of such quantification measurements.

Animals↗
PubMed2005

Accuracy of liver fat quantification at MR imaging: comparison of out-of-phase gradient-echo and fat-saturated fast spin-echo techniques--initial experience.

PURPOSE: To retrospectively determine the relative accuracy of liver fat quantification with out-of-phase gradient-echo magnetic resonance (MR) imaging and fat-saturated fast spin-echo MR imaging in patients with and without cirrhosis, with histologic analysis as the reference standard. MATERIALS AND METHODS: Committee on Human Research approval was obtained. Patient consent was not required. Data collection ended before HIPAA regulations were implemented, but patient anonymity was maintained. Twenty-seven patients, 16 with cirrhosis, were retrospectively identified who underwent MR imaging before histopathologic evaluation of liver fat at biopsy or surgery. The patient population consisted of 15 male and 12 female patients (mean age, 55 years; range, 16-75 years). One radiologist blinded to the histopathologic results recorded mean signal intensity derived from three regions of interest placed in the right and left lobes of the liver on three sections and signal intensity of the spleen from one region of interest within the same section. Liver fat was quantified with the relative loss of signal intensity on out-of-phase images compared with that on in-phase T1-weighted gradient-echo images and with relative loss of signal intensity on T2-weighted fast spin-echo MR images obtained with fat saturation compared with those obtained without fat saturation. Hotelling t test was used to compare correlation coefficients between relative signal intensity differences and histopathologically determined percentage of fat. RESULTS: In patients without cirrhosis, liver fat quantification with fat-saturated fast spin-echo MR imaging was significantly better than it was with out-of-phase gradient-echo MR imaging (r = 0.92 vs 0.69, P < .01). In patients with cirrhosis, liver fat quantification was correlated only with fat-saturated fast spin-echo MR imaging (r = 0.76, P < .01); the relative signal intensity loss on out-of-phase gradient-echo MR images was not correlated with histopathologically determined percentage of fat (r = 0.25, P = .36). CONCLUSION: Preliminary results suggest liver fat may be more accurately quantified with fat-saturated fast spin-echo MR imaging than with out-of-phase gradient-echo MR imaging, especially in patients with cirrhosis.

Adolescent↗
PubMed2006

1H MR spectroscopy of the brain: absolute quantification of metabolites.

Hydrogen 1 (1H) magnetic resonance (MR) spectroscopy enables noninvasive in vivo quantification of metabolite concentrations in the brain. Currently, metabolite concentrations are most often presented as ratios (eg, relative to creatine) rather than as absolute concentrations. Despite the success of this approach, it has recently been suggested that relative quantification may introduce substantial errors and can lead to misinterpretation of spectral data and to erroneous metabolite values. The present review discusses relevant methods to obtain absolute metabolite concentrations with a clinical MR system by using single-voxel spectroscopy or chemical shift imaging. Important methodological aspects in an absolute quantification strategy are addressed, including radiofrequency coil properties, calibration procedures, spectral fitting methods, cerebrospinal fluid content correction, macromolecule suppression, and spectral editing. Techniques to obtain absolute concentrations are now available and can be successfully applied in clinical practice. Although the present review is focused on 1H MR spectroscopy of the brain, a large part of the methodology described can be applied to other tissues as well.

Brain↗
PubMed1994

Hemochromatosis: diagnosis and quantification of liver iron with gradient-echo MR imaging.

PURPOSE: To assess the role of magnetic resonance (MR) imaging in detection and quantification of liver iron overload. MATERIALS AND METHODS: MR imaging at 0.5 T was prospectively performed on 77 patients (67 with liver iron overload and 10 without) who underwent a liver biopsy with biochemical determination of the liver iron concentration (LIC) (normal, < 36 mumol per gram of liver tissue [dry weight]). Ratios of signal intensities and liver T2 relaxation time were calculated from images obtained with spin-echo and breath-hold gradient-echo (GRE) sequences. RESULTS: Liver-to-tissue signal intensity ratios were better correlated with LIC than T2 relaxation time. Long-echo-time GRE sequences were the most sensitive for detection of slight overload. Thus, high sensitivity (94%) and specificity (90%) were obtained with a liver-to-fat ratio threshold of 1. The quantification of iron with MR imaging was accurate when the LIC was 80-300 mumol/g. For heavy overload, above 300 mumol/g, quantification was impossible owing to complete signal loss. Pancreatic and splenic signal intensity were unchanged in most cases. CONCLUSION: This method, which can be improved by using more sensitive sequences with a high-field-strength system, should be competitive with biopsy for the diagnosis of substantial liver iron overload.

Female↗
PubMed1995

Carotid artery stenosis: clinical efficacy of MR phase-contrast flow quantification as an adjunct to MR angiography.

PURPOSE: To assess the utility of magnetic resonance (MR) flow quantification in the evaluation of suspected carotid artery stenosis. MATERIALS AND METHODS: Fifty-five patients referred for angiography and 10 healthy volunteers underwent Doppler ultrasound, three-dimensional time-of-flight MR angiography, and MR phase-contrast flow quantification to measure peak systolic velocity (PSV) and volumetric flow rate (VFR) in the common and internal carotid arteries distal to the stenosis. RESULTS: PSV and VFR were significantly lower in the severely (> or = 70%) stenosed internal carotid arteries (P < .05). The VFR ratio (internal carotid artery-common carotid artery) achieved 91% overall accuracy for detection of severe stenosis. Combination of flow data with results of MR angiography yielded sensitivity of 100% (95% confidence interval, 78%-100%) with a modest loss in specificity. Doppler and MR measurements of PSV in the common carotid artery showed significant correlation: volunteers, r = .73; patients, r = .64. CONCLUSION: MR flow quantification provides information about the hemodynamic significance of carotid stenosis. As an adjunct to MR angiography, it may be useful in enabling differentiation of occlusions from critical stenoses.

Adult↗
PubMed1998

Automatic quantification of immunohistochemically stained cell nuclei based on standard reference cells.

A fully automatic method for quantification of images of immunohistochemically stained cell nuclei by computing area proportions, is presented. Agarose embedded cultured fibroblasts were fixed, paraffin embedded and sectioned at 4 microm. They were then stained together with 4 microm sections of the test specimen obtained from bladder cancer material. A colour based classifier is automatically computed from the control cells. The method was tested on formalin fixed paraffin embedded tissue section material, stained with monoclonal antibodies against the Ki67 antigen and cyclin A protein. Ki67 staining results in a detailed nuclear texture with pronounced nucleoli and cyclin A staining is obtained in a more homogeneously distributed pattern. However, different staining patterns did not seem to influence labelling index quantification, and the sensitivity to variations in light conditions and choice of areas within the control population was low. Thus, the technique represents a robust and reproducible quantification method. In tests measuring proportions of stained area an average standard deviation of about 1.5% for the same field was achieved when classified with classifiers created from different control samples.

Automation↗
PubMed2004

Quantification of colorectal cancer micrometastases in lymph nodes by nested and real-time reverse transcriptase-PCR analysis for carcinoembryonic antigen.

PURPOSE: Reverse-transcriptase PCR (RT-PCR) assays for carcinoembryonic antigen (CEA) have been described to identify lymph node micrometastases. These assays are not quantitative and can be confounded by false-positive results. The purpose of this study was to determine whether quantification of CEA in lymph nodes could more readily identify clinically relevant groups. EXPERIMENTAL DESIGN: Specimens included 400 lymph nodes from 64 patients undergoing colon resections. Specimens were tested by immunohistochemistry and by RT-PCR using nested primers for CEA. Specimens from 59 patients that were positive by nested RT-PCR were further quantified by detection of CEA mRNA fluorescence increase at a threshold PCR cycle. RESULTS: CEA was detected by nested RT-PCR analysis in 4 of 34 (12%) nodes of nonneoplastic disease, 2 of 13 (15%) nodes from T(1)N(0) patients, 32 of 81 (40%) nodes of T(2)N(0) patients, 49 of 109 (45%) nodes from T(3)N0 patients, and 92 of 163 (56%) nodes from T(1-4)N(1-2) patients. The overall presence of any RT-PCR-detectable CEA in nodes did not differentiate patient groups. Immunohistochemistry was positive in nodes from 7% of T(3)N(0) patients and 100% of T(1-3)N(1-2) patients. CEA quantification revealed that 0 of 7 patients with nonneoplastic disease and 2 of 17 (12%) patients with stage I T(1-2)N(0) cancers had one or more lymph nodes with >/=1.0 x 10(2) CEA transcripts per sample. In contrast, 4 of 13 (31%) patients with stage II T(3)N(0) cancer and 10 of 22 (45%) stage III patients with known metastases had lymph nodes with >/=1.0 x 10(2) CEA transcripts. CONCLUSIONS: These data suggest that quantification of CEA levels in lymph nodes may more accurately identify patients at risk for cancer recurrence than does routine nested RT-PCR or immunohistochemistry.

Carcinoembryonic Antigen↗
PubMed1993

Detection and quantification of calcium fluoride using micro-Raman spectroscopy.

Investigations on the caries-preventive effect of calcium fluoride have been seriously hampered by the lack of adequate detection techniques. In this paper, the micro-Raman technique is introduced as a suitable method for CaF2 quantification with a spot size typically 5 microns. Advantages of this measuring technique and associated problems are described for CaF2 determination in the presence of large amounts of hydroxyapatite. The data show that a comparative analysis of Raman intensities of calcium fluoride and phosphate ions of hydroxyapatite can be used for quantification of CaF2 concentrations. The intensity behaviour of the CaF2 and hydroxyapatite Raman lines is explained considering the absorption-scattering cross-sections of the two components. The ratio of the Raman intensity (322 cm-1) of CaF2 to the phosphate Raman intensity (432/447 cm-1) of hydroxyapatite was found to be related linearly to the ratio of the concentration of CaF2 to that of hydroxyapatite. At present, the lower limit of CaF2 quantification is approximately 3 wt%. The technique has already been applied on fluoridated bovine enamel, and Raman emissions due to CaF2 or CaF2-like material formed on/in enamel have been detected for the first time.

Animals↗