Simultaneous Analysis of Multiple Chromosomal Aberrations in Chronic Lymphocytic Leukemia (CLL) by Multiplex Ligation-Dependent Probe Amplification (MLPA).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2329-2329
Author(s):  
Marian J.P.L. Stevens-Kroef ◽  
Annet Simons ◽  
Michael Egmont Petersen ◽  
Hanneke Gorissen ◽  
Yvonne Nanciu ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
False Negative ◽  
Lymphocytic Leukemia ◽  
Prognostic Information ◽  
Interphase Fish ◽  
Negative Results ◽  
False Negative Results ◽  
Trisomy 12 ◽  
Abnormal Cells ◽  
Dependent Probe

Abstract B cell chronic lymphocytic leukemia (B-CLL) is characterized by a highly variable clinical course. Characteristic genomic abnormalities have shown to provide highly significant prognostic information (Dohner NEJM 2000). However, karyotyping and/or FISH are laborious and expensive techniques. Although other prognostic markers (like mutation status of VH-genes and expression of CD38 and ZAP70) have been noted, these fail to identify all patients with a cytogenetic high-risk profile. We have performed multiplex ligation-dependent probe amplification (MLPA) analysis on 37 CLL patients, using the P037 and P038 kits (mrc-holland, http://www.mlpa.com/pages/indexpag.html), which allow the simultaneous identification of 56 genomic CLL-specific targets. The MLPA data obtained were compared with interphase FISH data in a blinded fashion using the ATM, centromere 12, RB1, D13S319, and TP53 specific probes (Vysis). By doing so, a good correlation was revealed in those cases in which the percentage of abnormal cells was at least 27%. We diagnosed six cases with loss of the ATM locus, six cases with trisomy 12, twenty cases with loss of chromosome 13q14 (three of them with a biallelic deletion), and three cases with loss of the TP53 locus. In three cases the genetic abnormality was detected by interphase FISH only. In these latter cases, the percentages of abnormal cells were 13%, 20% and 20%, respectively, which may explain the false negative results of the MLPA analyses. Since multiple probes were used for the 13q14 loci, a critical region of genomic loss could be identified which excluded the RB1 gene but included the KCNRG, DLEU2 and DLEU1 genes and miRNA15A. Also for the TP53 and ATM genes multiple probes were tested and, based on these results, one patient is suspected to have an intragenic ATM deletion. Also, additional abnormalities were detected by MLPA as e.g. trisomy 19. In conclusion, MLPA appears to be a useful technique for the identification of clinically relevant genetic abnormalities in a diagnostic setting.

Chronic Lymphocytic Leukemia, Detection of Five Significant Prognostic Genomic Aberrations by Interphase Two-Color Fluorescence in Situ Hybridization Analyses: a Study of 24 Patients with Clinical Follow-up Ranged From 7 to 120 Months.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4716-4716
Author(s):  
Carlos Sergio Barragan ◽  
Silvia B. Hansson ◽  
Silvina A Cicchini ◽  
Monica Tamashiro ◽  
Pablo G Dinardo ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chronic Lymphocytic Leukemia ◽  
Fluorescence In Situ Hybridization ◽  
Lymphocytic Leukemia ◽  
The Other ◽  
Prognostic Information ◽  
Trisomy 12 ◽  
Genomic Aberrations

Abstract Abstract 4716 Background Conventional cytogenetic studies by chromosome banding are difficult in chronic lymphocytic leukemia (CLL) because of the low in vitro proliferation or mitotic activity of CLL cells. Interphase Fluorescence in situ hybridization (FISH) has improved the detection of genomic aberrations in CLL. We used this method to identify chromosomal abnormalities in patients with CLL at diagnosis, progression disease, after relapse or treatment failure. Methods Mononuclear cells from the blood of 24 Caucasian patients, 12 men and 12 women, age ranged from 39 to 84 years (median, 64) with CLL were analyzed by FISH, for deletions in chromosome bands 11q22.3, 6q23.3, 13q14.3, 17p13.1, and trisomy of chromosome 12. The patients stages were 0A:12; IA:1; IIA: 2; IIB:4; IIIA:1, IIIB:1; IIIC:1; IV:2. The follow up ranged was from 7 to 124 months. Results Chromosomal aberrations were detected in 15 of 24 (58.33 percent). The most frequent changes were a 12 trisomy (37.5 percent), a deletion in 11q23.3, (12.5 percent), a deletion in 6q23.3 (8.33 percent), a deletion in 17p13.1 ( 8.33 percent), a deletion in 13q14.3 8.33%. A patient showed four aberrations tri(12), d(17p13.1), d(6q23.3), d(11q23.3), she was resistant to Fludarabine + Cyclophosfamide treatment and Rituximab CHOP and had a survival of 51 months. The other patients with trisomy 12 alone are still alive and the survival were ranged from 7 to 120 month. Trisomy 12 was correlated with enlarged lymph nodes. Two patients undergo Richter transformation one of them with 6q23.3 the other without the aberrations assessed. The patients with deletion 11q23.3 were resistant to standard regimens treatment. Ten patients have stable disease (0A), clinical follow up ranged from 7 to 124 months; three with trisomy 12 the others without the genetic aberrations assesses. Longest survivals (124 month) were found in patients without aberration or with trisomy 12. Patients without aberrations or with trisomy 12 had similar evolution. A female patient, 84 years old state in stage 0A along 13 month with d(17p13.1), and d(11q23.3). Patients with 11q23.3 deletions had the shortest median treatment-free interval (11months), and those with trisomy 12 or deletion 13q14.3 had the longest (124 and 40 months respectively). Patients with 11q23.3 and 6q23.3 deletion had more advanced disease. Two patients died, both of them with 6q23.3, overall survival 51 and 54 months respectively. ZAP 70 and CD38 do not have correlation with presence of the genomic aberration assessed .The present of 11q23.3, 6q23.3, 17p13.1, age, the white-cell count, Rai - Binet stage and the serum lactate dehydrogenase level gave significant prognostic information. Conclusions Genomic aberrations are independent prognosis factors of disease progression and survival in Chronic lymphocytic leukemia; establish them properly will be an important goal standard, therefore incorporate FISH assay routinely could give significant prognostic information. These genetic findings are predictors of outcome treatment regimens and could help to choose the appropriated strategic treatments. Disclosures: No relevant conflicts of interest to declare.

Critical Evaluation of Impedance Phlebography for the Diagnosis of Deep Vein Thrombosis

1974 ◽  
Vol 31 (02) ◽  
pp. 273-278
Author(s):  
Kenneth K Wu ◽  
John C Hoak ◽  
Robert W Barnes ◽  
Stuart L Frankel
Keyword(s):  
Deep Vein Thrombosis ◽  
False Positive ◽  
False Negative ◽  
Critical Evaluation ◽  
Original Method ◽  
Vein Thrombosis ◽  
Negative Results ◽  
False Negative Results ◽  
Deep Vein ◽  
Positive Results

SummaryIn order to evaluate its daily variability and reliability, impedance phlebography was performed daily or on alternate days on 61 patients with deep vein thrombosis, of whom 47 also had 125I-fibrinogen uptake tests and 22 had radiographic venography. The results showed that impedance phlebography was highly variable and poorly reliable. False positive results were noted in 8 limbs (18%) and false negative results in 3 limbs (7%). Despite its being simple, rapid and noninvasive, its clinical usefulness is doubtful when performed according to the original method.

THE ROLE OF THE GENETIC ABNORMALITIES, EPIGENETIC AND microRNA IN THE PROGNOSIS OF CHRONIC LYMPHOCYTIC LEUKEMIA

2018 ◽  
Vol 40 (4) ◽  
pp. 261-267 ◽  
Author(s):  
K Tari ◽  
Z Shamsi ◽  
H Reza Ghafari ◽  
A Atashi ◽  
M Shahjahani ◽  
...  
Keyword(s):  
B Cells ◽  
Chronic Lymphocytic Leukemia ◽  
Kinase Inhibitors ◽  
Bone Marrow Involvement ◽  
P53 Mutation ◽  
Lymphocytic Leukemia ◽  
Gene Promoters ◽  
Epigenetic Changes ◽  
Genetic Changes ◽  
Trisomy 12

Chronic lymphocytic leukemia (CLL) is increased proliferation of B-cells with peripheral blood and bone marrow involvement, which is usually observed in older people. Genetic mutations, epigenetic changes and miRs play a role in CLL pathogenesis. Del 11q, del l17q, del 6q, trisomy 12, p53 and IgVH mutations are the most important genetic changes in CLL. Deletion of miR-15a and miR-16a can increase bcl2 gene expression, miR-29 and miR-181 deletions decrease the expression of TCL1, and miR-146a deletion prevents tumor metastasis. Epigenetic changes such as hypo- and hypermethylation, ubiquitination, hypo- and hyperacetylation of gene promoters involved in CLL pathogenesis can also play a role in CLL. Expression of CD38 and ZAP70, presence or absence of mutation in IgVH and P53 mutation are among the factors involved in CLL prognosis. Use of monoclonal antibodies against surface markers of B-cells like anti-CD20 as well as tyrosine kinase inhibitors are the most important therapeutic approaches for CLL.

Scoring System for the Diagnosis of COVID-19

2020 ◽  
Vol 13 (1) ◽  
pp. 413-414 ◽  
Author(s):  
Mohamed Farouk Allam
Keyword(s):  
Scoring System ◽  
Clinical Symptoms ◽  
False Negative ◽  
Nasopharyngeal Swab ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Negative Results ◽  
False Negative Results ◽  
Symptoms And Signs

Due to the international spread of COVID-19, the difficulty of collecting nasopharyngeal swab specimen from all suspected patients, the costs of RT-PCR and CT, and the false negative results of RT-PCR assay in 41% of COVID-19 patients, a scoring system is needed to classify the suspected patients in order to determine the need for follow-up, home isolation, quarantine or the conduction of further investigations. A scoring system is proposed as a diagnostic tool for suspected patients. It includes Epidemiological Evidence of Exposure, Clinical Symptoms and Signs, and Investigations (if available). This scoring system is simple, could be calculated in a few minutes, and incorporates the main possible data/findings of any patient.

Detection of Streptococcus pneumoniae in Sputum Samples by Real- Time PCR.

2020 ◽  
Vol 18 ◽  
Author(s):  
Pegah Shakib ◽  
Mohammad Reza Zolfaghari
Keyword(s):  
Streptococcus Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
False Negative ◽  
Cross Sectional Study ◽  
Laboratory Culture ◽  
Cross Sectional ◽  
Negative Results ◽  
False Negative Results ◽  
Pcr Method

Background: Conventional laboratory culture-based methods for diagnosis of Streptococcus pneumoniae are time-consuming and yield false negative results. Molecular methods including real-time (RT)-PCR rapid methods and conventional PCR due to higher sensitivity and accuracy have been replaced instead traditional culture assay. The aim of the current study was to evaluate lytA gene for detection of Streptococcus pneumoniae in the cerebrospinal fluid of human patients with meningitis using real-time PCR assay. Material and Methods: In this cross-sectional study, a total of 30 clinical specimens were collected from patients in a period from September to December 2018. In order to evaluate the presence of lytA gene, conventional and real-time PCR methods were used without culture. Results: From 30 sputum samples five (16.66%) isolates were identified as S. pneumoniae by lytA PCR and sequencing. Discussion: In this research, an accurate and rapid real-time PCR method was used, which is based on lytA gene for diagnosis of bacteria so that it can be diagnosed. Based on the sequencing results, the sensitivity for detection of lytA gene was 100% (5/5).

COVID-19: how nuclear medicine can provide a differential diagnosis in very dubious case

Coronaviruses ◽  
2020 ◽  
Vol 01 ◽  
Author(s):  
Maria Silvia De Feo ◽  
Viviana Frantellizzi ◽  
Giuseppe De Vincentis
Keyword(s):  
Infectious Disease ◽  
Differential Diagnosis ◽  
Ct Scan ◽  
Imaging Technique ◽  
False Negative ◽  
Clinical Signs ◽  
Negative Results ◽  
Infectious Disease Department ◽  
False Negative Results ◽  
99Mtc Hmpao

Background: We present the case of a 55-year-old woman, admitted to the Infectious Disease Department of Policlinico Umberto I, Rome, in mid-March 2020, with suspicion of COVID-19 infection. Objective: The rRT-PCR was negative and the following CT scan, performed to exclude false-negative results and help diagnosis, was inconclusive. Methods: It was decided to submit the patient to 99mTc-HMPAO-labelled leukocyte scan. Results: This exam led to the diagnosis of infective endocarditis. Conclusion: In the present pandemic scenario, 99mTc-HMPAO-labelled leukocyte scan represents a reliable imaging technique for differential diagnosis with COVID-19 in patients with confusing clinical signs, possible false-negative rRT-PCR results and inconclusive CT scan.

Aspiration Revisited: Prospective Evaluation of a Physiologically Pressurized Model With Animal Correlation and Broader Applicability to Filler Complications

2021 ◽  
Author(s):  
Hyoung-Jin Moon ◽  
Won Lee ◽  
Ji-Soo Kim ◽  
Eun-Jung Yang ◽  
Hema Sundaram
Keyword(s):  
Normal Saline ◽  
False Negative ◽  
Vascular Complications ◽  
Filler Injection ◽  
Negative Results ◽  
Needle Position ◽  
False Negative Results ◽  
In Vivo Testing

Abstract Background Aspiration testing before filler injection is controversial. Some believe that aspiration can help prevent inadvertent intravascular injection, while others cite false-negative results and question its value given that the needle position always changes somewhat during injection procedures. Objectives To test the relation of false-negative results to the viscosity of the material within the needle lumen and determine whether a less viscous material within the needle lumen could decrease the incidence of false-negative results. Methods In vitro aspiration tests were performed using 30-G and 27-G needle gauges, two cross-linked hyaluronic acid fillers, normal saline bags pressurized at 140 and 10 mmHg to mimic human arterial and venous pressures, and three needle lumen conditions (normal saline, air, and filler). Testing was repeated three times under each study condition (72 tests in total). For in vivo correlation, aspiration tests were performed on femoral arteries and central auricular veins in three rabbits (4–5 aspirations per site, 48 tests in total). Results In vitro and in vivo testing using 30-G needles containing filler both showed false-negative results on aspiration testing. In vitro and in vivo testing using needles containing saline or air showed positive findings. Conclusions False-negative results from aspiration testing may be reduced by pre-filling the needle lumen with saline rather than a filler. The pressurized system may help overcome challenges of animal models with intravascular pressures significantly different from those of humans. The adaptability of this system to mimic various vessel pressures may facilitate physiologically relevant studies of vascular complications.

scholarly journals Superiority of MALDI-TOF Mass Spectrometry over Real-Time PCR for SARS-CoV-2 RNA Detection

Viruses ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 730
Author(s):  
Magda Rybicka ◽  
Ewa Miłosz ◽  
Krzysztof Piotr Bielawski
Keyword(s):  
Mass Spectrometry ◽  
Gold Standard ◽  
Viral Genome ◽  
False Negative ◽  
Rt Pcr ◽  
Rna Detection ◽  
Maldi Tof ◽  
Negative Results ◽  
False Negative Results ◽  
Pcr Test

At present, the RT-PCR test remains the gold standard for early diagnosis of SARS-CoV-2. Nevertheless, there is growing evidence demonstrating that this technique may generate false-negative results. Here, we aimed to compare the new mass spectrometry-based assay MassARRAY® SARS-CoV-2 Panel with the RT-PCR diagnostic test approved for clinical use. The study group consisted of 168 suspected patients with symptoms of a respiratory infection. After simultaneous analysis by RT-PCR and mass spectrometry methods, we obtained discordant results for 17 samples (10.12%). Within fifteen samples officially reported as presumptive positive, 13 were positive according to the MS-based assay. Moreover, four samples reported by the officially approved RT-PCR as negative were positive in at least one MS assay. We have successfully demonstrated superior sensitivity of the MS-based assay in SARS-CoV-2 detection, showing that MALDI-TOF MS seems to be ideal for the detection as well as discrimination of mutations within the viral genome.

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