Prognostic Value of Circulating Melanoma Cells Detected by Reverse Transcriptase–Polymerase Chain Reaction

2003 ◽  
Vol 21 (5) ◽  
pp. 767-773 ◽  
Author(s):  
Giuseppe Palmieri ◽  
Paolo A. Ascierto ◽  
Francesco Perrone ◽  
Sabrina M.R. Satriano ◽  
Alessandro Ottaiano ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Subgroup Analysis ◽  
Peripheral Blood ◽  
Predictive Value ◽  
Prognostic Value ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Blood Samples ◽  
Stage Of Disease ◽  
Polymerase Chain

Purpose: Factors that are predictive of prognosis in patients who are diagnosed with malignant melanoma (MM) are widely awaited. Detection of circulating melanoma cells (CMCs) by reverse transcriptase-polymerase chain reaction (RT-PCR) has recently been postulated as a possible negative prognostic factor. Two main questions were addressed: first, whether the presence of CMCs, defined as the patient being positive for any of the three markers, had a prognostic role; and second, what the predictive value of each individual marker was. Patients and Methods: A consecutive series of 200 melanoma patients observed between January 1997 and December 1997, with stage of disease ranging from I to IV, was analyzed by semiquantitative RT-PCR. Tyrosinase, p97, and MelanA/MART1 were used as markers to CMCs on baseline peripheral blood samples. Progression-free survival (PFS) was used as a unique end point and was described by the product limit method. Multivariable analysis was applied to verify whether the auspicated prognostic value of these markers was independent of the stage of disease, and a subgroup analysis was performed that excluded patients with stage IV disease. Results: Overall, 32% (64 of 200) of patients progressed, and a median PFS of 52 months in the whole series was observed. The presence of CMCs and the markers individually or combined was predictive of prognosis in the univariate analysis but did not provide additional prognostic information to the stage of disease in multivariable models. In the subgroup analysis of stage (ie, I–III subgroup), similar results were observed. Conclusion: Detection of CMCs in peripheral blood samples at the time of MM diagnosis by semiquantitative RT-PCR does not add any significant predictive value to the stage of disease. Thus, this approach should not be used in clinical practice, and further studies are required to determine its usefulness.

Impact of Molecular Staging Methods in Primary Melanoma: Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) of Ultrasound-Guided Aspirate of the Sentinel Node Does Not Improve Diagnostic Accuracy, But RT-PCR of Peripheral Blood Does Predict Survival

2008 ◽  
Vol 26 (35) ◽  
pp. 5742-5747 ◽  
Author(s):  
Christiane A. Voit ◽  
Gregor Schäfer-Hesterberg ◽  
Martina Kron ◽  
Alexander C.J. van Akkooi ◽  
Juergen Rademaker ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Peripheral Blood ◽  
Primary Melanoma ◽  
Disease Free Survival ◽  
Rt Pcr ◽  
Ultrasound Guided ◽  
Chain Reaction ◽  
Blood Samples ◽  
Polymerase Chain

Purpose This study analyzes (1) the value of tyrosinase reverse-transcriptase polymerase chain reaction (RT-PCR) of aspirates obtained by ultrasound-guided fine-needle aspiration cytology (US-FNAC) of sentinel nodes (SNs) in patients with melanoma before sentinel lymph node biopsy (SLNB) and (2) the value of RT-PCR of blood samples of all SLNB patients. Patients and Methods Between 2001 and 2003, 127 patients with melanoma (median Breslow depth, 2.1 mm) underwent SLNB. FNAC was performed in all SNs of all patients pre- and post-SLNB. The aspirates were partly shock-frozen for RT-PCR and were partly used for standard cytology. Peripheral blood was collected at the time of SLNB and at every outpatient visit thereafter. Results Thirty-four (23%) of 120 SNs were positive for melanoma. SN involvement was predicted by US-FNAC with a sensitivity of 82% and a specificity of 72%. Additional tyrosinase RT-PCR revealed the same sensitivity of 82% and a specificity of 72%. At a median follow-up time of 40 months from first blood sample, peripheral-blood RT-PCR was a significant independent predictor of disease-free survival (DFS) and overall survival (OS; P < .001). Conclusion US-FNAC is highly accurate and eliminates the need for SLNB in 16% of all SLNB patients. RT-PCR of the aspirate or excised SN does not improve sensitivity or specificity. RT-PCR of blood samples predicts DFS and OS.

GeneXpert Essay for BCR-ABL Compared to RT-PCR for Clinical Management of CML

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4416-4416
Author(s):  
G. Pica ◽  
G. Catania ◽  
F. Castelli ◽  
C Repetto ◽  
A. M. Carella ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Standard Deviation ◽  
Predictive Value ◽  
Gene Fusion ◽  
Negative Control ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Prolymphocytic Leukemia ◽  
Blood Samples ◽  
Polymerase Chain

Abstract Abstract 4416 Reverse transcriptase-polymerase chain reaction (RT-PCR) is considered the most sensitive method available for detecting low copy numbers of the BCR-ABL gene fusion. The difference between the BCR-ABL Ct (threshold cycle) and ABL Ct is expected to represent the ratio of the two populations of mRNAs and ultimately the percentage of neoplastic cells present. An international scale (IS) has been proposed for BCR-ABL RQ-PCR measurements. To enable testing centers to gain access to the IS, the Adelaide laboratory initiated a process to develop and validate laboratory-specific conversion factors (CFs) that can be used to convert local values to IS values. Recently, Cepheid introduced its GeneXpert-based assay for the identification of the BCR-ABL gene fusion in cells from blood samples. This system comprises a walkaway self-contained instrument that combines cartridge- based microfluidic sample preparation with reverse transcriptase-polymerase chain reaction-based fluorescent signal detection and BCR-ABL and ABL Ct determination. The CF provided by Cepheid for this procedure is 0.47. We tested this BCR-ABL fusion detection system, compared with a classical RT-PCR analysis as a clinical diagnostic tool for CML patients. We tested 19 patient peripheral blood samples by both methods. The negative control group included 3 blood samples, obtained from 3 patients with hematological disorders unrelated to BCR-ABL gene fusion: one with essential thrombocythemia, one with chronic myelomonocytic leukemia, one with T-cell-prolymphocytic leukemia. The remaining 16 clinical samples belonged to 12 patients with an established diagnosis of CML in chronic phase and to one patient with Philadelphia chromosome-positive acute lymphoblastic leukemia (p210). GeneXpert software reported as negative 2 samples with a numerical threshold limit calculated on Ct (i.e. BCR-ABL was not detected at a detection limit of 0.00046%); the 3rd negative control was indicated as BCR-ABL invalid: this result was probably due to the high proportion of ABL detected as consequence of hyperleukocytosis (WBC 559,6 × 109/L) in T-Cell prolymphocytic leukemia patients. Therefore, the performance of GeneXpert test in this series was: Sensitivity 100%, Specificity 0,66%, Positive Predictive Value 100%, Negative Predictive Value 100%. Among the series of 16 true positive samples GeneXpert revealed mean 8,67; mean standard deviation 27,55; median 1,48. Whereas in the same series RT-PCR results were: mean 37,09; mean standard deviation 59,33; median 3,37. In conclusion GeneXpert essay for BCR-ABL could be an important tool for diagnose and monitor CML but require complete clinical data for the correct interpretation of results. Disclosures: No relevant conflicts of interest to declare.

scholarly journals A meta-analysis of accuracy and sensitivity of chest CT and RT-PCR in COVID-19 diagnosis

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Fatemeh Khatami ◽  
Mohammad Saatchi ◽  
Seyed Saeed Tamehri Zadeh ◽  
Zahra Sadat Aghamir ◽  
Alireza Namazi Shabestari ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Ct Scan ◽  
Predictive Value ◽  
Meta Analysis ◽  
Chest Ct ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Chest Ct Scan

AbstractNowadays there is an ongoing acute respiratory outbreak caused by the novel highly contagious coronavirus (COVID-19). The diagnostic protocol is based on quantitative reverse-transcription polymerase chain reaction (RT-PCR) and chests CT scan, with uncertain accuracy. This meta-analysis study determines the diagnostic value of an initial chest CT scan in patients with COVID-19 infection in comparison with RT-PCR. Three main databases; PubMed (MEDLINE), Scopus, and EMBASE were systematically searched for all published literature from January 1st, 2019, to the 21st May 2020 with the keywords "COVID19 virus", "2019 novel coronavirus", "Wuhan coronavirus", "2019-nCoV", "X-Ray Computed Tomography", "Polymerase Chain Reaction", "Reverse Transcriptase PCR", and "PCR Reverse Transcriptase". All relevant case-series, cross-sectional, and cohort studies were selected. Data extraction and analysis were performed using STATA v.14.0SE (College Station, TX, USA) and RevMan 5. Among 1022 articles, 60 studies were eligible for totalizing 5744 patients. The overall sensitivity, specificity, positive predictive value, and negative predictive value of chest CT scan compared to RT-PCR were 87% (95% CI 85–90%), 46% (95% CI 29–63%), 69% (95% CI 56–72%), and 89% (95% CI 82–96%), respectively. It is important to rely on the repeated RT-PCR three times to give 99% accuracy, especially in negative samples. Regarding the overall diagnostic sensitivity of 87% for chest CT, the RT-PCR testing is essential and should be repeated to escape misdiagnosis.

Detection of circulating neoplastic cells by reverse-transcriptase polymerase chain reaction in malignant melanoma: association with clinical stage and prognosis.

1996 ◽  
Vol 14 (7) ◽  
pp. 2091-2097 ◽  
Author(s):  
B Mellado ◽  
D Colomer ◽  
T Castel ◽  
M Muñoz ◽  
E Carballo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Malignant Melanoma ◽  
Prognostic Factor ◽  
Peripheral Blood ◽  
Clinical Stage ◽  
Melanoma Cells ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Circulating Neoplastic Cells

PURPOSE Circulating melanoma cells can be detected in peripheral blood by means of tyrosinase mRNA amplification by reverse-transcriptase polymerase chain reaction (RT-PCR). We conducted a prospective study to evaluate the clinical significance of the presence of circulating neoplastic cells in the blood of patients with malignant melanoma (MM). METHODS A sensitive RT-PCR assay was used to detect tyrosinase mRNA in the peripheral blood of patients with stages I to IV melanoma. Healthy subjects or patients with other malignancies were used as negative controls. RESULTS Ninety-one assessable patients were included in the study. There was a statistically significant association between RT-PCR positivity and clinical stage. Circulating melanoma cells were detected in 36% of patients with localized disease (stages I and II), in 45% of patients with regional nodal involvement (stage III), and in 94% of patients with metastatic disease (stage IV) (P < .001). In stage II-III patients who were RT-PCR-positive for mRNA tyrosinase in blood, the recurrence rate and disease-free survival were significantly worse than patients who were RT-PCR-negative. In multivariate analysis, RT-PCR was an independent prognostic factor for recurrence in patients with nonmetastatic disease (P = .002). CONCLUSION The detection of circulating melanoma cells in peripheral blood by RT-PCR correlated with the clinical stage of patients with melanoma and was an independent prognostic factor for recurrence. Further studies are warranted to better assess the significance of this test in the evaluation of prognosis, early detection of relapse, and in monitoring the effectiveness of systemic therapy.

Polymerase Chain Reaction-Based Detection of Circulating Melanoma Cells as an Effective Marker of Tumor Progression

1999 ◽  
Vol 17 (1) ◽  
pp. 304-304 ◽  
Author(s):  
Giuseppe Palmieri ◽  
Maria Strazzullo ◽  
Paolo A. Ascierto ◽  
Sabrina M.R. Satriano ◽  
Antonio Daponte ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Tumor Progression ◽  
Peripheral Blood ◽  
Melanoma Cells ◽  
Disease Stage ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Gene Markers ◽  
Risk Of Recurrence ◽  
Polymerase Chain

PURPOSE: Reverse transcriptase (RT) polymerase chain reaction (PCR) with multiple markers has been demonstrated to be highly sensitive in detecting circulating cells from patients with malignant melanoma (MM). We evaluated the clinical significance of the presence in peripheral blood of specific PCR-positive mRNA markers as an expression of circulating melanoma cells. PATIENTS AND METHODS: Total cellular RNA was obtained from the peripheral blood of 235 patients with either localized (n = 154) or metastatic (n = 81) melanoma. We performed RT-PCR using tyrosinase, p97, MUC18, and MelanA/MART1 as gene markers. The PCR products were analyzed by gel electrophoresis and Southern blot hybridization. In addition, 20 healthy subjects and 21 patients with nonmelanoma cancer were used as negative controls. RESULTS: Although detected at various levels among assessable patients, each mRNA marker was significantly correlated with disease stage. A significant correlation with disease stage was demonstrated for patients who were positive to all four markers (P < .0001) or to at least three markers (P < .001). Univariate analysis showed a significant correlation between risk of recurrence (evaluated in stage I, II, and III patients) and increasing number of PCR-positive markers (P = .0002). Logistic regression multivariate analysis indicated that each single marker (except tyrosinase) and, more especially, the presence of four PCR-positive markers remained statistically independent prognostic factors for tumor progression. CONCLUSION: Our data establish the existence of a significant correlation among clinical stages, tumor progression, and presence of circulating melanoma-associated antigens in peripheral blood of MM patients. Preliminary assessment of a subset of patients with a higher risk of recurrence needs longer follow-up and further studies to define the role of RT-PCR in monitoring MM patients.

scholarly journals The utility of real-time polymerase chain reaction in detecting Coccidioides immitis among clinical specimens in the Central California San Joaquin Valley

2018 ◽  
Vol 57 (6) ◽  
pp. 688-693 ◽  
Author(s):  
Dominic Dizon ◽  
Marilyn Mitchell ◽  
Bernadette Dizon ◽  
Robert Libke ◽  
Michael W Peterson
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Pleural Fluid ◽  
Clinical Setting ◽  
Predictive Value ◽  
Coccidioides Immitis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Acute Pneumonia ◽  
Polymerase Chain

AbstractCoccidioidomycosis, the fungal infection caused by dimorphic Coccidioides species, is typically diagnosed by histopathologic identification of spherules, by culture, or by serology. These tests are reliable but time-intensive, delaying diagnosis and treatment. Rapid real-time polymerase chain reaction (RT-PCR) can be performed and was validated to identify Coccidioides immitis using an in-house developed assay for the Becton Dickinson molecular instrument (BD MAXTM). These studies were performed using patient samples that had been shown to be positive on previously set up fungal cultures. To evaluate this new RT-PCR test in the clinical setting, we conducted a retrospective chart review of patients (N = 1160) who underwent Coccidioides PCR (Cocci PCR) on clinical samples between March 1, 2014, and Dec 31, 2016. We abstracted clinical, microbiologic, serologic, radiographic, treatment, and follow-up data. Specimens of cerebrospinal fluid (CSF), bronchioalveolar lavage fluid (BAL), lung tissue biopsy (LTB), sputum, and pleural fluid were evaluated to determine sensitivity and specificity. Of the 113 specimens that tested positive for Cocci PCR, all had clinical disease defined by traditional clinical criteria, yielding 100% specificity. Overall sensitivity was 74% versus 46% for fungal culture and was available in 4 hours rather than 1–2 weeks. Sensitivities varied by source material and clinical setting. CSF had a sensitivity of 59%, BAL for acute pneumonia 91%, sputum for acute pneumonia 94%, pleural fluid 86%, but LTB for lung nodules only 44%. Overall positive predictive value (PPV) was 100%, while negative predictive value (NPV) was 96%, but again this varied by specimen and clinical setting. Our experience with clinical testing of >1160 specimens over 2–3 years shows we can utilize this technology to improve our ability to diagnose disease but that the sensitivity varies by specimen source and clinical setting.

Prospective Multi-Institutional Study of Reverse Transcriptase Polymerase Chain Reaction for Molecular Staging of Melanoma

2006 ◽  
Vol 24 (18) ◽  
pp. 2849-2857 ◽  
Author(s):  
Charles R. Scoggins ◽  
Merrick I. Ross ◽  
Douglas S. Reintgen ◽  
R. Dirk Noyes ◽  
James S. Goydos ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Peripheral Blood ◽  
Prognostic Significance ◽  
Disease Free Survival ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Molecular Staging ◽  
Polymerase Chain

Purpose To evaluate the prognostic significance of molecular staging using reverse transcriptase polymerase chain reaction (RT-PCR) in detecting occult melanoma cells in sentinel lymph nodes (SLNs) and circulating bloodstream. Patients and Methods In this multicenter study, eligibility criteria included patient age 18 to 71 years, invasive melanoma ≥ 1.0 mm Breslow thickness, and no clinical evidence of metastasis. SLN biopsy and wide excision of the primary tumor were performed. SLNs were examined by serial-section histopathology and S-100 immunohistochemistry. A portion of each SLN was frozen for RT-PCR. In addition, RT-PCR was performed on peripheral-blood mononuclear cells (PBMCs). RT-PCR analysis was performed using four markers: tyrosinase, MART1, MAGE3, and GP-100. Disease-free survival (DFS), distant–DFS (DDFS), and overall survival (OS) were analyzed. Results A total of 1,446 patients with histologically negative SLNs underwent RT-PCR analysis. At a median follow-up of 30 months, there was no difference in DFS, DDFS, or OS between the RT-PCR–positive (n = 620) and RT-PCR–negative (n = 826) patients. Analysis of PBMC from 820 patients revealed significant differences in DFS and DDFS, but not OS, for patients with detection of more than one RT-PCR marker in peripheral blood. Conclusion In this large, prospective, multi-institutional study, RT-PCR analysis on SLNs and PBMCs provides no additional prognostic information beyond standard histopathologic analysis of SLNs. Detection of more than one marker in PBMC is associated with a worse prognosis. RT-PCR remains investigational and should not be used to direct adjuvant therapy at this time.

scholarly journals Is the Portable Chest Radiographic More Reliable to Reveal Covid19 in Highly Suspicion Patient before Real-Time Reverse Transcription Polymerase Chain Reaction (RT-PCR) Test?

2021 ◽  
Vol 6 (03) ◽  
pp. 228-236
Author(s):  
Bushra A. A. Albazi ◽  
Dr Noof. Albaz ◽  
Dr Nayef. Alqahtani ◽  
Dr. Angham Salih ◽  
Dr Rafat Mohtasab
Keyword(s):  
Polymerase Chain Reaction ◽  
Retrospective Study ◽  
Predictive Value ◽  
Rt Pcr ◽  
Chain Reaction ◽  
X Ray ◽  
Polymerase Chain ◽  
Rate Type ◽  
Test Result ◽  
Pcr Test

A large number of patients with coronavirus disease 2019 (COVID-19) present at hospitals. There are a limited number of isolation rooms open, and patients must often wait a long time to get a reverse transcription-polymerase chain reaction (RT-PCR) test done. This necessitates the introduction of effective triage plans. A patient with suspicions is referred to an emergency room (ED) depending on their medical record for a simple physical assessment, blood test findings, and chest imaging.A retrospective study design was conduct at Prince Sultan Medical Military City (PSMMC). Ethical approval was obtained from the institutional board to wave the consent forms since it is a retrospective study. Only the primary investigator has had the data access to the patients’ medical records. The collected patient records were under specific categories, including symptoms score starts from 5 and above, RT-PCR test result done after CXRP imaging, the patient admitted to the emergency department (ED). Excluding all CXRP done after RT-PCR TEST, positive Covid 19 admitted to the intensive care unit (ICU), pediatric patients, and patients with score symptoms were less than five. Two experienced radiologists reviewed the images blindly, and the inter-observer reliability of observations noted by the radiologists was calculated. As for the relationship between the x-ray reading and the RT-PCR test result, our results showed a high correlation between the variables (chi-square χ² = 12.44, with df =1, and p<0.001). The sensitivity of x-ray diagnosing covid19 was 65.52 %, while the specificity was 54.51 %, and the accuracy of radiologists reading was 58.17 %. Furthermore, the positive predictive value (PPV) was 41.76 %, and the negative predictive value (NPV) was 76.05%. Finally, the false positive rate (type-i error (alpha) was 45.49%, and the false-negative rate (type-ii error (beta) was 34.48% Our research findings show that CXRP imaging can detect COVID-19 infection in symptomatic patients and can be a valuable addition to RT-PCR testing. In an inpatient ED environment where availability of test kits, laboratory equipment, and laboratory personnel is compromised and risks delaying patient treatment and hospital workflow, serial CXRP could theoretically be used as an adjunct diagnostic function and monitoring in patients suspected of having COVID-19.

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