Discovery of molecular biomarker signatures for the detection of bladder cancer.

302 Background: Bladder cancer (BCa) is among the five most common malignancies world-wide, and due to high rates of recurrence, one of the most prevalent. Improvements in non-invasive urine-based assays to detect BCa would benefit both patients and healthcare systems. In this study, the goal was to identify urothelial cell transcriptomic signatures associated with BCa. Methods: Gene expression profiling (Affymetrix U133 Plus 2.0 arrays) was applied to exfoliated urothelia obtained from a cohort of 92 subjects with known bladder disease status. Computational analyses identified candidate biomarkers of BCa and an optimal predictive model was derived. Selected targets from the profiling analyses were monitored in an independent cohort of 81 subjects using quantitative real-time PCR (RT-PCR). Results: Data analysis identified 52 genes associated with BCa (p≤0.001), and gene models that optimally predicted class label were derived. RT-PCR analysis of 48 selected targets in an independent cohort identified a 14-gene diagnostic signature that predicted the presence of BCa with a specificity of 100% at 90% sensitivity. Conclusions: Exfoliated urothelia sampling provides a robust analyte for the evaluation of patients with suspected BCa. The refinement and validation of the multi-gene urothelial cell signatures identified in this preliminary study may lead to accurate, non-invasive assays for the detection of BCa. The development of an accurate, non-invasive BCa detection assay would benefit both the patient and healthcare systems through better detection, monitoring and control of disease.

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Rapid detection of SARS-CoV-2 infection by multicapillary column coupled ion mobility spectrometry (MCC-IMS) of breath. A proof of concept study

10.1101/2020.06.30.20143347 ◽

2020 ◽

Cited By ~ 2

Author(s):

Claus Steppert ◽

Isabel Steppert ◽

Gunther Becher ◽

William Sterlacci ◽

Thomas Bollinger

Keyword(s):

Discriminant Analysis ◽

Ion Mobility ◽

Influenza A ◽

Cross Validation ◽

Viral Disease ◽

Pcr Analysis ◽

Proof Of Concept ◽

Influenza Epidemic ◽

Rt Pcr ◽

Non Invasive

AbstractThere is an urgent need for screening patients of having a communicable viral disease to cut infection chains.We could recently demonstrate that MCC-IMS of breath is able to identify Influenza-A infected patients. With decreasing Influenza epidemic and upcoming SARS-CoV-2 infections we extended our study to the analysis of patients with suspected SARS-CoV-2 infections.51 patients, 23m, 28f, aged 64 ± 16 years, were included in this study.Besides RT-PCR analysis of nasopharyngeal swabs all patients underwent MCC-IMS analysis of breath. 16 patients, 7m, 9f, were positive for SARS-CoV-2 by RT-PCR. There was no difference in gender or age according to the groups.Stepwise canonical discriminant analysis could correctly classify the infected and non-infected subjects in 98% by cross-validation. Afterwards we combined the Influenza-A sub study and the SARS-CoV-2-sub study for a total of 75 patients, 34m, 41f, aged 64.8 ± 1.8 years, 14 positive for Influenza-A, 16 positive for SARS-CoV-2, the remaining 44 patients were used as controls. In one patient RT-PCR was highly suspicious of SARS-CoV-2 but inconclusive.There was no imbalance between the groups for age or gender.97.3% of the patients could be correctly classified to the respective group by discriminant analysis. Even the inconclusive patient could be mapped to the SARS-CoV-2 group applying the discrimination function.ConclusionMCC-IMS is able to detect SARS-CoV-2 infection and Influenza-A infection in breath. As this method provides exact, fast non-invasive diagnosis it should be further developed for screening of communicable viral diseases.Study registration: NCT04282135

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Rapid detection of SARS-CoV-2 infection by multicapillary column coupled ion mobility spectrometry (MCC-IMS) of breath. A proof of concept study

10.21203/rs.3.rs-70139/v1 ◽

2020 ◽

Author(s):

Claus Steppert ◽

Isabel Steppert ◽

William Sterlacci ◽

Thomas Bollinger

Keyword(s):

Ion Mobility ◽

Influenza A ◽

Viral Disease ◽

Pcr Analysis ◽

Proof Of Concept ◽

Influenza Epidemic ◽

Rt Pcr ◽

Respective Group ◽

Non Invasive ◽

Multicapillary Column

Abstract There is an urgent need for screening of patients having a communicable viral disease to cut infection chains. We could recently demonstrate that MCC-IMS of breath is able to identify Influenza-A infected patients. With decreasing Influenza epidemic and upcoming SARS-CoV-2 infections we went on and also analysed patients with suspected SARS-CoV-2 infections.75 patients, 34m, 41f, aged 64.4 ± 15.4 years, 14 positive for Influenza-A, 16 positive for SARS-CoV-2, the remaining 44 patients were used as controls. In one patient RT-PCR was highly suspicious of SARS-CoV-2 but initially inconclusiveBesides RT-PCR analysis of nasopharyngeal swabs all patients underwent MCC-IMS analysis of breath. There was no difference in gender or age according to the groups.97.3% of the patients could be correctly classified to the respective group by discriminant analysis. Even the inconclusive patient could be mapped to the SARS-CoV-2 group applying the discrimination function.ConclusionMCC-IMS is able to detect SARS-CoV-2 infection and Influenza-A infection in breath. As this method provides exact, fast non-invasive diagnosis it should be further developed for screening of communicable viral diseases.Trial registrationClinicalTrial.gov, NCT04282135 Registered 20 February 2020 - Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT04282135?term=IMS&draw=2&rank=1

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Molecular detection of Mycobacterium tuberculosis in pulmonary and extrapulmonary samples in a hospital-based study

African Health Sciences ◽

10.4314/ahs.v20i4.14 ◽

2020 ◽

Vol 20 (4) ◽

pp. 1617-23

Author(s):

Kalal Iravathy Goud ◽

Matam Kavitha ◽

Adi Mahalakshmi ◽

Ravi Vempati ◽

Abdulaziz A Alodhayani ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Pcr Analysis ◽

Sequencing Analysis ◽

Rt Pcr ◽

Tissue Samples ◽

Non Invasive ◽

Invasive Test ◽

Keywords Tuberculosis ◽

Bronchoalveolar Fluid

Objective: Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a deadly infectious disease. India contributes to one-third of the global TB burden. However, no studies have been carried out in the Telangana (Hyderabad) population using real-time polymerase chain reaction (RT-PCR). Therefore, the current study evaluated the role of RT-PCR as a rapid and non-invasive test to diagnose TB by testing for pulmonary tuberculosis (PTB) and extrapulmonary tubercu- losis (EPTB). Materials and methods: This hospital-based study examined 1670 samples (900 EPTB; 770 PTB) comprising tissue (n = 537), peritoneal fluid (n = 420), sputum (n = 166), bronchial fluid (n = 126), cerebrospinal fluid (n = 145), ascetic fluid (n = 76), sputum pus (n = 78), urine (n = 79), and bronchoalveolar fluid (n = 43) samples. DNA from samples was separated using specific isolation kits and subjected to RT-PCR. Results: In this study, we enrolled 1670 subjects and categorized 54.4% as females and 45.6% as males. The collected sam- ples were categorized as 48.5% of fluid samples, followed by tissue (32.2%), sputum (9.9%), urine (4.7%), and pus-swab (4.6%). RT-PCR analysis revealed that 4.7% patients were positive for Mtb. Our results revealed that 61% of the affected patients were male and 39% were female. Among the specimen types, tissue samples gave the highest proportion of positive results (36.3%). Conclusion: The results showed that RT-PCR should be implemented and given top priority in TB diagnosis to save time and facilitate a definitive diagnosis. Tissue samples are highly recommended to screen the Mtb through the technique RT- PCR. Future studies should extend the technique to the global population and exome sequencing analysis should be per- formed to identify TB risk markers. Keywords: Tuberculosis (TB); EPTB; PTB; Mycobacterium tuberculosis (Mtb).

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Tumor and Microenvironment Modification during Progression of Murine Orthotopic Bladder Cancer

Clinical and Developmental Immunology ◽

10.1155/2011/865684 ◽

2011 ◽

Vol 2011 ◽

pp. 1-11 ◽

Cited By ~ 5

Author(s):

Sin Mun Tham ◽

Kee Hui Ng ◽

Sim Hwee Pook ◽

Kesavan Esuvaranathan ◽

Ratha Mahendran

Keyword(s):

Gene Expression ◽

Bladder Cancer ◽

Gene Expression Analysis ◽

Therapeutic Strategies ◽

Pcr Analysis ◽

Low Density ◽

Rt Pcr ◽

Tumor Implantation ◽

C3h Mice ◽

Immune Related Genes

The aim of this study was to monitor changes in the expression of immune-related genes in the bladder after tumor implantation. Mice were orthotopically implanted with MB49-PSA cells (C57BL/6 mice) on day 1 and terminated on days 7, 14, 21, and 28. Another mouse model (MBT-2/C3H mice) was examined at day 7. Gene expression analysis was performed using a TaqMan Low Density Mouse Immune Panel (Applied Biosystems, USA) on RNA extracted from the bladders. Selected genes were reconfirmed by real-time PCR analysis and RT-PCR on the mRNA from other animals. Immune suppressive (IL13, IL1β, PTGS2, NOS2, IL10, CTLA4, and CCL22) and immune stimulatory genes (CSF2, GZMB, IFNγ, CXCL10, TNFα, CD80, IL12a, and IL6) and AGTR2 were increased by day 7. By day 28, IL10, CCL2, CCL5, CXCL11, CTLA4, GZMB, IFNγ, CSF2, and IL6 were significantly increased. Therapeutic strategies involving TH1 induction and TH2 dampening may improve responses to immunotherapy.

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Does sampling saliva increase detection of SARS-CoV-2 by RT-PCR? Comparing saliva with oro-nasopharyngeal swabs

10.1101/2020.07.26.20158618 ◽

2020 ◽

Cited By ~ 1

Author(s):

Ozlem Akgun Dogan ◽

Betsi Kose ◽

Nihat Bugra Agaoglu ◽

Jale Yildiz ◽

Gizem Alkurt ◽

...

Keyword(s):

Gold Standard ◽

Viral Rna ◽

Pcr Analysis ◽

Rt Pcr ◽

Standard Samples ◽

Rna Detection ◽

Detection Rates ◽

Gold Standard Method ◽

Non Invasive ◽

Isolation Period

The gold standard method in the diagnosis of SARS-CoV-2 infection is the detection of viral RNA in nasopharyngeal sample by RT-PCR. Recently, saliva samples has been suggested as an alternative due to being fast, reliable and non-invasive, rather than nasopharyngeal samples. We compared RT-PCR results in nasopharyngeal, oro-nasopharyngeal and saliva samples of COVID-19 patients. 98 of 200 patients were positive in RT-PCR analysis performed before the hospitalization. In day 0, at least one sample was positive in 67% of 98 patients. Positivity rate was 83% for both oro-nasopharyngeal and nasopharyngeal samples, while it was 63% for saliva samples (p<0.001). On day 5, RT-PCR was performed in 59 patients, 34% had at least one positive result. The positivity rate was 55% for saliva and nasopharyngeal samples, while it was 60% for oro-nasopharyngeal samples. Our study shows that the sampling saliva does not increase the sensitivity of RT-PCR tests at early stages of infection. However, on 5th day, viral RNA detection rates in saliva were similar to nasopharyngeal and oro-nasopharyngeal samples. In conclusion, we suggest that, in patients receiving treatment, virus presence in saliva, in addition to the standard samples, is important to determine the isolation period and to control the transmission.

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Identification of miR-29c-3p as a Robust Normalizer for Urine microRNA Studies in Bladder Cancer

Biomedicines ◽

10.3390/biomedicines8110447 ◽

2020 ◽

Vol 8 (11) ◽

pp. 447

Author(s):

Julia Oto ◽

Emma Plana ◽

Álvaro Fernández-Pardo ◽

Fernando Cana ◽

Manuel Martínez-Sarmiento ◽

...

Keyword(s):

Bladder Cancer ◽

Diagnostic Marker ◽

Quantitative Polymerase Chain Reaction ◽

Diagnostic Techniques ◽

Expression Study ◽

Non Invasive ◽

Good Expression ◽

Polymerase Chain ◽

Early Diagnose ◽

Independent Cohort

Bladder cancer (BC) is among the most frequent malignancies worldwide, being the most expensive cancer to treat and monitor and the most lethal urological cancer. Urine microRNAs (miRNAs) have been proposed as novel non-invasive biomarkers to early diagnose and monitor BC patients in order to avoid the performance of current aggressive diagnostic techniques. However, huge discrepancies arise among studies mainly due to the lack of standardization in the normalization, a crucial step in all miRNA studies. Our aim was to identify the best miRNA normalizer for miRNA studies in urine of BC patients. We evaluated the performance of 110 candidate miRNAs in urine of 35 BC patients and 15 healthy controls by Real Time quantitative Polymerase Chain Reaction (RT-qPCR) followed by a stability analysis with RefFinder. In this screening stage, miR-29c-3p arose as the most stably expressed miRNA in BC and controls, with a good expression level. Stability of miR-29c-3p expression was validated in an independent cohort of 153 BC patients and 57 controls. Finally, we evaluated the robustness of miR-29c-3p as normalizer in the expression study of miR-200c-3p, a potential diagnostic marker for BC. We propose miR-29c-3p as a normalizer for miRNA studies in BC urine. This is the first study that characterizes a reliable normalizer that may allow the comparison of future urine miRNA studies as non-invasive biomarkers for BC diagnosis and monitoring.

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Non-invasive urine based tests for the detection of bladder cancer

Journal of Clinical Pathology ◽

10.1136/jclinpath-2012-200812 ◽

2012 ◽

Vol 65 (11) ◽

pp. 970-975 ◽

Cited By ~ 18

Author(s):

Neha Wadhwa ◽

Suresh Kumar Jatawa ◽

Archana Tiwari

Keyword(s):

Bladder Cancer ◽

New Technologies ◽

Recent Literature ◽

Malignant Neoplasm ◽

Healthcare Systems ◽

Women Patients ◽

Urinary Markers ◽

Non Invasive ◽

Current Review ◽

Higher Sensitivity

Bladder cancer is the fourth most frequently diagnosed malignant neoplasm and cause of cancer-related deaths in men and eighth in women. Patients with bladder cancer undergo repeated cystoscopic examinations of the bladder to monitor for tumour recurrence which is invasive, costly and lacks accuracy. Therefore, the development of non-invasive urine based tests for the early detection of bladder cancer would be of tremendous benefit to both patients and healthcare systems. A number of urine based markers are available for the early diagnosis of bladder cancer. The diagnosis of bladder cancer relies on identifying malignant cells in the urine. All urinary markers have a higher sensitivity as compared with cytology but they score lower in specificity. Many soluble and cell based markers have been developed. Only two of the soluble and cell based markers have obtained the Food and Drug Administration approval. In the current review, the most recent literature of urinary markers is summarised. This article reports some of the more prominent urine markers and new technologies used nowadays.

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