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Reduction of false-positive results with biochemical second-tier testing for newborn screening of Pompe disease.

PURPOSE: To review the performance and outcomes of a second-tier newborn screening test for Pompe disease. METHODS: We followed our previously published screening approach that reduces false-positive results by incorporating creatine and creatinine levels and postanalytic tools in a second-tier test. RESULTS: We reviewed 1879 blood samples from neonates born in 11 states. Second-tier testing effectively reduced false-positive results, compared with first-tier enzyme testing alone. Only a small number of screen-positive cases (n = 7) were confirmed to have infantile-onset Pompe disease. No false-negative cases of infantile-onset Pompe disease were identified in this cohort, and 6 cases of possible late-onset Pompe disease were not detected with this approach. CONCLUSION: This tiered screening strategy discriminated well between true- and false-positive results and improved the positive predictive value. However, it did not reliably differentiate between infantile- and late-onset Pompe disease.

Humans

Using Mapping-Profiles to Refine Strain-Level Metagenomic Classification.

Metagenomic classification at the strain level remains challenging due to high sequence similarity among closely related genomes, which leads to ambiguous read mappings and frequent false-positive strain detections. Reducing such errors improves the reliability of strain-level analyses, which is critical for applications such as pathogen detection. We introduce StrainRefine, a post-mapping refinement method that analyzes read-reference mapping profiles to resolve ambiguous assignments among highly similar genomes. The method represents candidate reference genomes using binary profiles that capture read-support patterns and measures similarity between references based on profile overlap. The method clusters references based on similar mapping profiles, filters weakly supported genomes, and reassigns reads to representative references, reducing redundant reporting of near-identical strains. StrainRefine substantially reduces false-positive strain detections while preserving recall and improving agreement between predicted and true abundance profiles. On large-scale metagenomic datasets, it achieves a substantially improved precision-recall balance compared with existing mapping-based approaches, with the standalone method obtaining the highest read-level classification accuracy on the most complex evaluated dataset. Unlike many strain-level tools designed for individual species, StrainRefine operates without prior assumptions about sample composition or curated species-specific reference collections, while still achieving comparable performance in single-species settings on species-specific reference databases. These results highlight mapping-profile similarity as an effective signal for improving strain-level metagenomic classification.

false-positive reduction

Rheumatoid factor interference with the latex agglutination test for fibrin degradation products.

The latex agglutination test for FDP is widely employed clinically to aid in the diagnosis of DIC and other conditions. Of sera containing RF, 93% demonstrated positive FDP latex agglutination tests. Reducing agents in all instances destroyed the RF agglutinating capability. Futhermore, 86% of sera positive for FDP and RF became FDP-negative following reduction. Therefore RF was responsible for false-positive FDP latex agglutination tests in the majority of patients. Reduction of patient sera is a rapid, simple method to distinguish a positive FDP test from a false-positive due to RF.

Arthritis

Noneffect of oral urinary copper ascorbic acid on reduction glucose test.

References and texts in the fields of diabetes and clinical chemistry commonly report that ascorbic acid when given orally or parenterally gives a false-positive reaction to the copper reduction glucose test (Clinitest). This impression is based on a study in which ascorbic acid (250 mg./dl.) was added to urine in vitro, with a resultant positive-test reading in the absence of glucose. Ascorbic acid is a reducing agent, and theoretically it could interfere with the copper reduction method of glucose detection. In the current study 10 nondiabetic men were ingesting 4 and 6 gm. ascorbic acid per day. A total of 360 glucose detection tests with the copper reduction method were undertaken. In no instance was there a positive reaction to the glucose test.

Administration, Oral

Noneffect of methyldopa on urine glucose tests.

Methyldopa has been reported to cause a false-positive urine glucose test by the copper reduction method (Clinitest). We investigated the effect of methyldopa on urine glucose tests in 30 patients by comparing commonly used glucose oxidase methods to Clinitest. The results of our study indicate that methyldopa does not interfere with the copper reduction method for urine glucose determination.

Copper

Evaluation of a caffeic acid-ferric citrate test for rapid identification of Cryptococcus neoformans.

An evaluation of a rapid caffeic acid-ferric citrate paper disk test for the identification of Cryptococcus neoformans, using 474 isolates of yeasts and yeastlike organisms, showed that 96.6, 97.7, and 98.3% of 176 isolates of C. neoformans produced brown to dark-brown pigment on disks incubated for 6 h at room temperature, 30 degrees C, and 37 degrees C, respectively. All C. neoformans produced brown to dark-brown pigment within 24 h. However, nonspecific pigmentation was encountered at all temperatures of incubation with one isolate of Trichosporon cutaneum and, at room temperature only, with one isolate of C. luteolus after 6 h. Other genera of yeasts produced similar pigmentation after 24 h at all temperatures. The age of the cultures tested or the types of media used before testing did not significantly affect the ability of C. neoformans to produce pigmentation at 37 degrees C. A positive test may prove useful for presumptive identification of C. neoformans, but a negative reaction should not be used to rule out an identification of this yeast. Since a number of false-negative and false-positive tests occur, it is necessary to confirm, by other biochemical tests, the identification of all organisms suspected of being C. neoformans, to reduct the serious risk of missing or misidentifying this important pathogen.

Caffeic Acids

Reduced anti-Mycobacterium tuberculosis antibody response in tuberculosis patients with acquired immunodeficiency syndrome.

1. When the sera of patients with tuberculosis were tested for anti-Mycobacterium tuberculosis IgG using an indirect ELISA, the test was positive for 94.1% of the samples from patients not having AIDS (N = 51), but for only 37.5% of the samples from patients with AIDS (N = 16). 2. False-positive results were obtained for 7.3% of patients not infected with HIV (N = 96) and for 4.7% of patients infected with HIV (N = 64). 3. In most serum samples obtained from patients with tuberculosis and AIDS after the beginning of specific treatment there was a reduction of the ELISA absorbance at 490 nm with time. 4. These results indicate that serological tests for the detection of anti-M. tuberculosis IgG in patients with AIDS are of limited value for the diagnosis of tuberculosis, most likely as a consequence of the underlying immune defect of the patients.

AIDS-Related Opportunistic Infections

Maternal serum screening for Down's syndrome: the effect of routine ultrasound scan determination of gestational age and adjustment for maternal weight.

OBJECTIVE: To investigate the effect of using a routine ultrasound estimate of gestational age and maternal weight adjustment on maternal serum alpha-fetoprotein (AFP), unconjugated oestriol (uE3) and human chorionic gonadotrophin (hCG) levels in antenatal screening for Down's syndrome. DESIGN: Women with a singleton pregnancy without Down's syndrome were screened using the three serum markers and an estimate of gestational age based on 'dates' (time since first day of the last menstrual period) and one based on an ultrasound scan examination was recorded together with maternal weight. SETTING: Women attending the Homerton Hospital, Hackney, for their antenatal care between February 1989 and January 1990. SUBJECTS: 2113 women with a singleton pregnancy without Down's syndrome. RESULTS: The use of ultrasound to estimate gestational age (usually based on the biparietal diameter of the fetal skull) led to a significant reduction in the variance of each marker at a given week of pregnancy. The level of each marker was negatively associated with maternal weight, so that adjustment for weight also led to a reduction in variance. These data on gestational age and maternal weight, taken together with published data on pregnancies associated with Down's syndrome, indicate that the routine use of ultrasound to estimate gestational age will increase the detection rate from 58% to 67% while maintaining the false-positive rate at 5%, or reduce the false-positive rate from 5.7% to 3.1% while maintaining the detection rate at 60%. Routine maternal weight adjustment for the serum marker levels was much less useful, increasing the detection rate by about 0.5% for a given false-positive rate, or reducing the false-positive rate about 0.1% for a given detection rate. CONCLUSION: An ultrasound gestational age estimate available at the time of Down's syndrome screening confers a substantial advantage to screening performance with a further small benefit resulting from maternal weight adjustment, which is worth adopting if it can be done without difficulty or extra cost.

Biomarkers

Maternal serum unconjugated oestriol and human chorionic gonadotrophin levels in pregnancies with insulin-dependent diabetes: implications for screening for Down's syndrome.

OBJECTIVE: To investigate maternal serum unconjugated oestriol (uE3) and human chorionic gonadotrophin (hCG) levels in pregnant women with insulin-dependent diabetes mellitus and to consider the implications of the results for antenatal screening for Down's syndrome. DESIGN: Descriptive study using stored antenatal serum samples. SETTING: Stored serum samples collected from women receiving routine antenatal care in Oxford. SUBJECTS: 126 singleton pregnancies in 92 women with insulin-dependent diabetes mellitus and for each pregnancy, two pregnancies without diabetes matched for gestational age and duration of storage of the serum sample. None of the pregnancies was associated with fetal neural tube defect or Down's syndrome. MAIN STUDY MEASURES: Maternal serum uE3 and hCG levels at 15-22 weeks gestation. Alpha-fetoprotein (AFP) levels were also measured for comparison. RESULTS: The median uE3 level in the diabetic pregnancies was 0.92 multiples of the median (MoM) for pregnancies without diabetes at the same gestational age (P less than 0.05); and the hCG level was 0.95 MoM (P = 0.48). The median AFP level was also reduced to 0.77 MoM (P less than 0.001). CONCLUSION: The reduction in uE3 and AFP levels in insulin-dependent diabetic pregnancies is sufficiently great to be taken into account in maternal serum screening programmes for Down's syndrome. Dividing the uE3 and AFP levels in such pregnancies by the corresponding median for insulin-dependent diabetic pregnancies will yield a similar false-positive rate in pregnancies with and without insulin-dependent diabetes mellitus.

Chorionic Gonadotropin

Myocsrdial uptake of technetium-99m stannous pyrophosphate following direct current transthoracic countershock.

The effect of direct current (DC) countershock upon myocardial technetium-99m stannous pyrophosphate (PYP) uptake was studied in 22 dogs. All eight dogs imaged had positive abnormal PYP scintigrams that were usually indistinguishable from experimental infarction. In three animals, additional areas of radionuclide uptake were seen in overlying noncardiac tissue. Left and right ventricular myocardial PYP uptake averaged (+/- SEM) 23 +/- 5 times control and 24 +/- 6 times control, respectively. These activity ratios occurred without reduction in regional myocardial blood flow (RMBF), and were associated with histologic evidence of necrosis. The necrosis was usually epicardial, corresponding to the transmural site of greatest PYP uptake. The magnitude of PYP accumulation and the weight of damaged tissue increased with increasing applied energy. Thus, PYP uptake following DC countershock could result in false-positive interpretation of acute ischemic myocardial infarction. Since RMBF is normal in regions of PYP uptake, the major determinant of radionuclide accumulation is the extent of cellular damage.

Animals

Peripheral pulmonary wedge angiography in chronic obstructive pulmonary disease. Relationship to pulmonary function, chest x-ray film, and hemodynamic data.

Peripheral pulmonary wedge angiographic studies can investigate only limited areas of the pulmonary vascular bed; nevertheless, this technique is very useful in emphasizing the morphologic changes of small vessels (less than 1 mm in diameter) in pulmonary emphysema. The aim of this work is to determine the relationship between peripheral wedge angiographic appearances and pulmonary function, the chest x-ray film, and hemodynamic data in patients with chronic obstructive pulmonary disease. Three healthy subjects, four patients with type-A chronic obstructive pulmonary disease, and eight patients with type-B chronic obstructive pulmonary disease were studied. Among the tests of pulmonary function, only a marked reduction (less than 60 percent of predicted) of the diffusing capacity for carbon monoxide is always indicative of severe widespread emphysema. No relationship was found between hemodynamic data and peripheral wedge angiographic findings. Some agreement resulted between the chest x-ray film and peripheral wedge angiographic abnormalities only in patients with type-A chronic obstructive pulmonary disease; on the contrary, in patients with type-B chronic obstructive pulmonary disease, both false-positives and false-negatives were noted.

Angiography

The spirographic "kink". A sign of emphysema.

The significance of the "kink" occurring in the early portion of the forced expiratory spirogram of some patients with obstructed airflow was investigated in 153 patients with chronic obstructive pulmonary disease or asthma. The kink, which is presumably due to airway collapse, is facilitated by the presence of emphysema, which results in loss of elastic recoil, a positive pleural pressure, high bronchiolar resistance, and structural weakness in the walls of the major airways. A significant reduction of carbon-monoxide diffusing capacity was used as the indicator of the presence of significant anatomic emphysema. Eighty percent (39) of 49 patients with the spirographic kink had a low diffusing capacity, whereas only 16 of those without the kink had significantly impaired diffusion. Seventy percent (39) of the 56 patients who had emphysema by this criterion demonstrated a spirographic kink. This test is, therefore, offered as a simple, effective, and widely applicable screening procedure for detecting emphysema, with a low incidence of false-positive results.

Asthma

An appraisal of the nitroblue tetrazolium reduction test.

The available literature on the nitroblue tetrazolium reduction test is reviewed. The mechanism of this test is poorly understood. There are two basic methods of performing the test, namely, the stimulated and the spontaneous methods. However, the test procedure has not been standardized, and numerous modifications have ensued resulting in a number of technics. The stimulated test has proved to be valuable for screening patients or carriers with chronic granulomatous disease of childhood. It is one of the useful tests in studying neutrophil dysfunction. On the other hand, extensive accumulated data indicate that this test, be it stimulted or spontaneous, cannot be relied upon as an absolute test to differentiate bacterial from nonbacterial infectious disease as there have been numerous false-positive and false-negative results. When this test is used, the result should be interpreted in the context of other laboratory test results and the clinical picture of the patient. Preliminary data indicate that in certain diseases, if the initial test is positive in a particular patient, it may be useful as one of the follow-up tests to ascertain the activity of the disease process.

Bacterial Infections

Role of 67gallium citrate scanning in the management of non-Hodgkin's lymphoma.

67Gallium scans were performed as part of the initial evaluation in 45 patients with non-Hodgkin's lymphoma. Eighteen of these patients underwent staging laparotomy and splenectomy. In addition, scans were performed either shortly after therapy was completed or during subsequent followup in 10 patients. The initial scans were found most useful for patients with histiocytic lymphoma: in detecting sites of involvement above the diaphragm and the high para-aortic/mesenteric region, and when tumors were greater than 2 cm in diameter. The addition of 67Ga scanning to the pre-operative clinical evaluation reduced the number of incorrectly staged patients from 8 to 4. Reversion of previously positive 67Ga scans to negative in 3 patients with suspected persistent or recurrent disease was associated with fibrosis and no lymphoma when biopsied. Five other patients had histologically documented positive 67Ga scans post-therapy; in 1 the 67Ga scan was only definitive noninvasive procedure. Despite the occurrence of both false-positive and false-negative 67Ga scans, this procedure appears to be a useful supplement to the pretreatment evaluation of patients with non-Hodgkin's lymphoma, especially the histiocytic form. Confirmation of its ability to detect high para-aortic/mesentric involvement may subsequently result in a reduction of the number of staging laparotomies necessary. For the post-treatment followup of these patients 67Ga scans may prove to be valuable noninvasive investigation.

Abdominal Neoplasms

A clinic-based system for monitoring the quality of techniques for the diagnosis of gonorrhea.

The author describes a clinic-based system that monitors the quality of techniques for diagnosis of gonorrhea by continuous comparison of the results of gram-strained smears of urethral and cervical specimens with the corresponding cultures. Discrepant results are immediately reviewed. During a two-year period, the system detected two defects in culture incubation, visual defects in a laboratory technician, and three unexplained reductions in cultural sensitivity. The system motivates clinicians and laboratory technicians to maintain a high level of quality control. In 11,474 comparisons of urethral smear and culture results, there was agreement in 96.9% while the smear alone was positive in 1.6% and the culture alone in 1.5%. Review of positive-smear-negative-culture discrepancies indicated that most smears judged "false-positive" were so classified on the basis of false-negative cultures. In study of 1,331 women, the endocervical smeras identified 60% (273/455) of those with positive culture discrepancies. Many of these may have represented false-negative cultures.

Clinical Laboratory Techniques

Results of ophthalmosonometry and supraorbital photoplethysmography in evaluating carotid arterial stenoses.

A total of 263 patients underwent either periorbital ophthalmosonometry (Doppler ultrasound) or a combination of ophthalmosonometry and supraorbital photoplethysmography (SOPPG) to determine the presence or absence of a hemodynamically significant lesion (occlusion or greater than 50% reduction in lumen diameter). There were 522 vessels studied with Doppler ultrasound and 272 vessels with a combination of the two modalities. The tests were correlated with carotid arteriography to determine their accuracy. Overall accuracy of the Doppler ultrasound examination was 95%. A normal Doppler ultrasound examination was obtained in 367 vessels and arteriography confirmed the absence of a hemodynamically significant lesion in 93%. Of 155 vessels with an abnormal Doppler examination, 149 (96%) were proven on arteriography to have hemodynamically significant lesions. The Doppler ultrasound examination had a high incidence of false-negative examinations (26%) in those vessels that were between 50% and 99% stenotic, however. The SOPPG alone had a high incidence of false-positive examinations (19%); however, in 155 vessels with a normal SOPPG, 98% were proved to have no hemodynamically significant lesion. The combination of Doppler ultrasound and SOPPG was highly accurate in identifying patients with hemodynamically significant lesions. Of 97 vessels with such lesions, 96 (99%) had either an abnormal Doppler ultrasound examination or an abnormal SOPPG. In 240 vessels with concordant results of the Doppler ultrasound and SOPPG, the presence or absence of a hemodynamically significant lesion was accurately predicted in 235 (98%).

Arterial Occlusive Diseases

Evaluation of the usefulness of nitroblue tetrazolium reduction test (NBT test) for detection of bacterial infection in cancer patients.

False positivity of the NBT test in neoplastic diseases described by some authose may be a limitation in the use of the test for infection screening in cancer patients. For explaining this problem studies were performed in a group of 30 patients with various untreated malignancies without infection and in 20 of these with bacteriologically confirmed infections. The obtained results were compared with those in groups of normal persons without and with infection. Slightly modified Park method for spontaneous and stimulated NBT test was used. No significant differences between patients of both groups without infection were found. The increase in positivity of the test in presence of infection was lower in cancer patients than in normal subjects, however, it was non-significant. We did not observe any false-positive results in our patients, though one case of false negativity in myeloma is described separately. It is concluded that neoplastic disease does not cause, as such positivity of the NBT test. Because of similar response of NBT test to presence of infection, the NBT test is recognized as very useful for infection screening in cancer patients similarly as in normal persons.

Adult