ABSTRACT
The rapid and presumptive detection of SARS-CoV-2 variants may be performed using multiplex RT-PCR assays. The aim of this study was to evaluate the diagnostic performance of five qualitative RT-PCR tests as compared with next-generation sequencing (NGS). We retrospectively examined a multi-variant panel (n = 72) of SARS-CoV-2-positive nasopharyngeal swabs categorized as variants of concern (Alpha, Beta, Gamma and Delta), variants under monitoring (Iota and Kappa) and wild-type strains circulating in Liguria (Italy) from January to August 2021. First, NGS libraries of study samples were prepared and mapped to the reference genome. Then, specimens were screened for the detection of L452R, W152C, K417T, K417N, E484Q, E484K and N501Y mutations using the SARS-CoV-2 Variants II Assay Allplex, UltraGene Assay SARS-CoV-2 452R & 484K & 484Q Mutations V1, COVID-19 Ultra Variant Catcher, SARS-CoV-2 Extended ELITe MGB and Simplexa SARS-CoV-2 Variants Direct. The overall accuracy of these assays ranged from 96.9% to 100%. Specificity and sensitivity were 100% and 96-100%, respectively. We highly recommend the use of these assays as second-level tests in the routine workflow of SARS-CoV-2 laboratory diagnostics, as they are accurate, user friendly, low cost, may identify specific mutations in about 2-3 h and, therefore, optimize the surveillance of SARS-CoV-2 variants.
ABSTRACT
BACKGROUND: In the literature, there exist conflicting data on the value of fibulin-3 (FBLN3) for the diagnosis of pleural effusion (PE) in malignant pleural mesothelioma (MPM). Therefore we compared the diagnostic performance of FBLN3 against that of soluble mesothelin-related peptide (SMRP) in a cohort of Italian patients. MATERIALS AND METHODS: FBLN3 and SMRP were detected in PE from 33 patients with MPM, 64 with pleural benign lesions and 23 with non-MPM pleural metastases using a commercial enzyme-linked-immunosorbent(ELISA)-assay kit according to manufacturers' instructions. RESULTS: Levels of FBLN3 were similar in PE from MPM and PE from other pathologies (geometric mean=68.1 vs. 66.2 ng/ml; p=0.872) in contrast to SMRP levels, which were significantly higher in PE from MPM (geometric mean=14.6 vs. 3.2 nM; p<0.001). Receiver operating characteristic analysis confirmed that SMRP showed a good performance (area under the curve=0.79, p<0.001), whereas FBLN3 was not able to discriminate MPM from other pathologies (area under the curve=0.44, p=0.838). CONCLUSION: FBLN3 detection in PE, in contrast to SMRP detection, is not useful as a biomarker for the diagnosis of PE from MPM.
Subject(s)
Extracellular Matrix Proteins/metabolism , Gene Expression Regulation, Neoplastic , Lung Neoplasms/metabolism , Mesothelioma/metabolism , Multidrug Resistance-Associated Proteins/metabolism , Pleural Effusion, Malignant/metabolism , Pleural Neoplasms/metabolism , Aged , Aged, 80 and over , Biomarkers, Tumor/metabolism , Cohort Studies , Enzyme-Linked Immunosorbent Assay , Female , Humans , Italy , Lung Neoplasms/pathology , Male , Mesothelioma/pathology , Mesothelioma, Malignant , Middle Aged , Pleural Effusion, Malignant/pathology , Pleural Neoplasms/pathology , ROC CurveABSTRACT
BACKGROUND/AIM: Mesothelin (SMRP) is regarded as a biomarker of malignant pleural mesothelioma (MPM). Herein, we analyzed the contribution of SMRP detection in pleural effusion and in serum to the diagnosis of MPM with non-positive cytology. MATERIALS AND METHODS: The present study included 52 cases of MPM, 43 of pleural benign lesions and 25 of non-MPM pleural metastases. SMRP was measured by MesoMark ELISA (Cis-Bio International Gif/Yvette; France). RESULTS: In non-positive cytology, effusion-SMRP showed higher diagnostic performance than serum-SMRP. We found 38 out of 52 (73.1%) cases of non-positive cytology MPM, out of which 27 (71.0%) were positive for effusion-SMRP (cut-off=12.70 nM) and 18 (47.4%) for serum-SMRP (cut-off=1.08 nM). When cytology, effusion- and serum-SMRP were used in combination, an overall sensitivity in detection of MPM of 78.9% was achieved. The same sensitivity was obtained by combining cytology with effusion-SMRP alone, whereas the combination of serum-SMRP with cytology led to a sensitivity of 61.5%. CONCLUSION: Detection of both effusion- and serum-SMRP can contribute to improve the diagnosis of MPM with non-positive cytology. However, the analysis of SMRP in effusion makes it unnecessary to test SMRP in the serum.
Subject(s)
Biomarkers, Tumor/blood , GPI-Linked Proteins/blood , Lung Neoplasms/diagnosis , Mesothelioma/diagnosis , Pleural Effusion/pathology , Pleural Neoplasms/diagnosis , Aged , Aged, 80 and over , Asbestosis/blood , Cytodiagnosis , Female , Humans , Lung Neoplasms/blood , Lung Neoplasms/pathology , Male , Mesothelin , Mesothelioma/blood , Mesothelioma/pathology , Mesothelioma, Malignant , Middle Aged , Neoplasm Metastasis , Pleural Neoplasms/blood , Pleural Neoplasms/pathologyABSTRACT
BACKGROUND/AIM: Soluble mesothelin-related peptide (SMRP) is regarded as a biomarker of malignant pleural mesothelioma (MPM). Herein, we compared the diagnostic performances of SMRP in matched pleural effusion (PE-SMRP) and serum (S-SMRP). MATERIALS AND METHODS: Diagnosis on pleural biopsies was performed for all patients including 43 with MPM, 23 with non-MPM pleural metastases (MTS) and 36 with benign (BNG) pleural diseases. SMRP was measured by a MesoMark ELISA (Cis-Bio International Gif/Yvette; France). RESULTS: Using the receiver operating characteristic (ROC) analysis, 12.70 and 1.08 nM were detected as cut-off values to optimal discrimination for PE-SMRP and S-SMRP, respectively. PE-SMRP showed a better diagnostic accuracy than S-SMRP in MPM vs. MTS+BNG (area under the ROC curve=81.6 vs. 70.5; sensitivity=69.8% vs. 46.5%; specificity=88.1% vs. 84.7%; diagnostic odds ratio (DOR)=17.1 vs. 4.8). In S-SMRP-negative patients, PE-SMRP maintained an acceptable performance (Sensitivity=47.8%; DOR=8.3; p=0.001), whereas in PE-SMRP-negative patients, S-SMRP performed very poorly (Sensitivity=15.4%; DOR=1.2; p=0.858). CONCLUSION: PE-SMRP detection has a superior diagnostic accuracy than S-SMRP detection in the diagnosis of MPM.
Subject(s)
GPI-Linked Proteins/analysis , GPI-Linked Proteins/blood , Lung Neoplasms/diagnosis , Mesothelioma/diagnosis , Aged , Aged, 80 and over , Biomarkers, Tumor/analysis , Biomarkers, Tumor/blood , Female , Humans , Lung Neoplasms/blood , Male , Mesothelin , Mesothelioma/blood , Mesothelioma, Malignant , Middle Aged , Neoplasm Metastasis/diagnosis , Odds Ratio , Pleural Effusion , Pleural Neoplasms/blood , Pleural Neoplasms/diagnosis , Prognosis , ROC Curve , ThoracoscopyABSTRACT
Soluble mesothelin-related peptide (SMRP) is regarded as an FDA approved biomarker for the diagnosis and monitoring of pleural malignant mesothelioma (MPM). We detected the SMRP levels in pleural effusions (PE) by means of an ELISA and analyzed their diagnostic relevance to differentiate MPM from benign pathology and from non-MPM pleural metastasis. Comparison with cytology in MPM-PE was also performed. We found that SMRP detection in MPM-PE can help the diagnosis of MPM and provide additional diagnostic value to cytology. We concluded that SMRP test may be incorporated into clinical practice of PE from patients suspicious for MPM.
Subject(s)
Biomarkers, Tumor/metabolism , GPI-Linked Proteins/metabolism , Mesothelioma/diagnosis , Pleural Effusion, Malignant/diagnosis , Pleural Neoplasms/diagnosis , Adult , Aged , Aged, 80 and over , Cytodiagnosis/methods , Enzyme-Linked Immunosorbent Assay/methods , Female , Humans , Male , Mesothelin , Mesothelioma/metabolism , Mesothelioma/pathology , Middle Aged , Peptides/metabolism , Pleural Effusion, Malignant/metabolism , Pleural Effusion, Malignant/pathology , Pleural Neoplasms/metabolism , Pleural Neoplasms/pathologyABSTRACT
Leptomeningeal (LM) carcinomatosis is an increasing clinical complication in patients with advanced breast cancer (BC). The LM carcinomatosis diagnostic procedures rely mainly on cerebrospinal fluid (CSF) cytology, although both the amount of CSF and the number of malignant cells remain limiting factors. Therefore, efforts should be made to design new highly sensitive diagnostic tools to detect malignant cells in CSF of BC patients with LM carcinomatosis. In this study, the human Mammaglobin (hMAM) mRNA amplification by RT-PCR was employed to detect metastatic cells in CSF and thus, to diagnose LM carcinomatosis in a BC patient. Our data demonstrate that hMAM transcripts are expressed in the CSF of a BC patient with LM carcinomatosis, hence making RT-PCR for hMAM a potentially suitable test to identify occult BC cells in the brain.
Subject(s)
Breast Neoplasms/cerebrospinal fluid , Meningeal Carcinomatosis/cerebrospinal fluid , Neoplasm Proteins/cerebrospinal fluid , Neoplasm Proteins/genetics , Uteroglobin/cerebrospinal fluid , Uteroglobin/genetics , Adult , Brain/metabolism , Breast Neoplasms/complications , Breast Neoplasms/pathology , Female , Humans , Mammaglobin A , Meningeal Carcinomatosis/complications , Meningeal Carcinomatosis/pathology , RNA, Messenger/metabolismABSTRACT
Human infection with the sheep nasal botfly Oestrus ovis is sporadic and is often the consequence of an accidental deposit of the larvae by an adult botfly in the eye. This infestation results in external ophthalmomyiasis that, although a very rare condition, is more common among people living close to farming communities. We report three cases of O. ovis infestation which occurred in Italy in a limited area of La Spezia province (Le Cinque Terre), Italy during summer 2004. None of the patients had contact with wild or farm animals.