Evaluation of a novel real-time PCR test based on the ssrA gene for the identification of group B streptococci in vaginal swabs

<p>Abstract</p> <p>Background</p> <p>Despite the implementation of prevention guidelines, early-onset group B streptococci (GBS) disease remains a cause of neonatal morbidity and mortality worldwide. Strategies to identify women who are at risk of transmitting GBS to their infant and the administration of intrapartum antibiotics have greatly reduced the incidence of neonatal GBS disease. However, there is a requirement for a rapid diagnostic test for GBS that can be carried out in a labour ward setting especially for women whose GBS colonisation status is unknown at the time of delivery. We report the design and evaluation of a real-time PCR test (<it>RiboSEQ </it>GBS test) for the identification of GBS in vaginal swabs from pregnant women.</p> <p>Methods</p> <p>The qualitative real-time PCR <it>RiboSEQ </it>GBS test was designed based on the bacterial <it>ssrA </it>gene and incorporates a competitive internal standard control. The analytical sensitivity of the test was established using crude lysate extracted from serial dilutions of overnight GBS culture using the IDI Lysis kit. Specificity studies were performed using DNA prepared from a panel of GBS strains, related streptococci and other species found in the genital tract environment. The <it>RiboSEQ </it>GBS test was evaluated on 159 vaginal swabs from pregnant women and compared with the GeneOhm™ StrepB Assay and culture for the identification of GBS.</p> <p>Results</p> <p>The <it>RiboSEQ </it>GBS test is specific and has an analytical sensitivity of 1-10 cell equivalents. The <it>RiboSEQ </it>GBS test was 96.4% sensitive and 95.8% specific compared to "gold standard" culture for the identification of GBS in vaginal swabs from pregnant women. In this study, the <it>RiboSEQ </it>GBS test performed slightly better than the commercial BD GeneOhm™ StrepB Assay which gave a sensitivity of 94.6% and a specificity of 89.6% compared to culture.</p> <p>Conclusion</p> <p>The <it>RiboSEQ </it>GBS test is a valuable method for the rapid, sensitive and specific detection of GBS in pregnant women. This study also validates the <it>ssrA </it>gene as a suitable and versatile target for nucleic acid-based diagnostic tests for bacterial pathogens.</p

Crop Updates 2003 - Geraldton

This session covers twenty eight papers from different authors Seasonal Outlook: What is in store for 2003, David Stephens, Department of Agriculture Examining The Management Options For Wheat Crops In The Coming Season, James Fisher, Department of Agriculture GMO’s – what do they offer? Ian Edwards, Grain Bio Tech Australia Pty Ltd The Big Gamble – Wheat prices for 2003, Dennis Wise, Profarmer Market outlook for other grains, Andrew Young, General Manager Agricorp Stripe rust – where to now for the WA wheat industry? Robert Loughman, Ciara Beard and Greg Shea, Department of Agriculture Baudin and Hamlin – new generation of malting barley developed in Western Australia, Blakely Paynter, Roslyn Jettner and Kevin Young, Department of Agriculture DBM in Canola, Kevin Walden, Department of Agriculture The latest on Lupin diseases, Geoff Thomas, Department of Agriculture Wheat variety performance in 2002 compared to the long term, Robin Wilson, Iain Barclay, Robyn McLean, Robert Loughman, Jenny Garlinge, Bill Lambe, Neil Venn and Peter Clarke, Department of Agriculture Do wide rows drought proof lupins on red loam? Martin Harries, Bob French, Wayne Parker and Murray Blyth, Department of Agriculture Do wide rows drought proof lupins on a sandy loam? Martin Harries, Bob French, Wayne Parker and Murray Blyth, Department of Agriculture Profit Proving Precision Agriculture, Peter Norris, Agronomy For Profit, Greg Lyle, CSIRO Land and Water, Yuna Farm Improvement Group Annual ryegrass seedbanks: the good, the bad, and the ugly, Kathryn Steadman, University of Western Australia, Amander Ellery, CSIRO Plant Industry, Sally C Peltzer, Department of Agriculture Wheat management packages for low rainfall areas, Kari-Lee Falconer, Department of Agriculture Ground water 1. Atrazine, Russell Speed, Department of Agriculture Groundwater 2. Current Trends, Russell Speed, Department of Agriculture Herbicide tolerance of wheat, lupins and pastures, Terry Piper and Harmohinder Dhammu, Department of Agriculture Farming with Tramlines, Bindi Webb, Paul Blackwell, Department of Agriculture, Phil Logue, Binnu, Nigel Moffat, Geraldton, Rohan Ford, Binnu, Miles Obst, Mingenew, The role of green manure crops in renovating poor performing paddocks: What’s it worth? Frances Hoyle, Leanne Schulz and Judith Devenish Department of Agriculture The looming threat of wild radish, Peter Newman, Department of Agriculture Does one ‘size’ fit all? Grant Morrow, Syngenta Crop Protection Climate Forecasts on the Internet, Ian Foster and David Stephens, Department of Agriculture Moisture delving = more reliable lupin establishment, Paul Blackwell, and Wayne Parker, Department of Agriculture Tramline Designs for better Weed control and Wheat value from non-spraying tramlines in a dry season, Paul Blackwell, Bindi Webb and Darshan Sharma, Department of Agriculture Biserrula Grazing Trial, Marnie Thomas, Department of Agriculture Performance of IT and TT canola varieties in the medium and high rainfall agzones of W.A., 2001-02, Graham Walton, Hasan Zaheer and Paul Carmody, Department of Agriculture Rapid Catchment Appraisal in Northern Agricultural Region, Mike Clarke, Paul Raper, Department of Agricultur

The impact of immediate breast reconstruction on the time to delivery of adjuvant therapy: the iBRA-2 study

Background: Immediate breast reconstruction (IBR) is routinely offered to improve quality-of-life for women requiring mastectomy, but there are concerns that more complex surgery may delay adjuvant oncological treatments and compromise long-term outcomes. High-quality evidence is lacking. The iBRA-2 study aimed to investigate the impact of IBR on time to adjuvant therapy. Methods: Consecutive women undergoing mastectomy ± IBR for breast cancer July–December, 2016 were included. Patient demographics, operative, oncological and complication data were collected. Time from last definitive cancer surgery to first adjuvant treatment for patients undergoing mastectomy ± IBR were compared and risk factors associated with delays explored. Results: A total of 2540 patients were recruited from 76 centres; 1008 (39.7%) underwent IBR (implant-only [n = 675, 26.6%]; pedicled flaps [n = 105,4.1%] and free-flaps [n = 228, 8.9%]). Complications requiring re-admission or re-operation were significantly more common in patients undergoing IBR than those receiving mastectomy. Adjuvant chemotherapy or radiotherapy was required by 1235 (48.6%) patients. No clinically significant differences were seen in time to adjuvant therapy between patient groups but major complications irrespective of surgery received were significantly associated with treatment delays. Conclusions: IBR does not result in clinically significant delays to adjuvant therapy, but post-operative complications are associated with treatment delays. Strategies to minimise complications, including careful patient selection, are required to improve outcomes for patients

Evaluation of a novel real-time pcr test based on the ssragene for the identification of group b streptococci in vaginal swabs

Background: Despite the implementation of prevention guidelines, early-onset group B streptococci (GBS) disease remains a cause of neonatal morbidity and mortality worldwide. Strategies to identify women who are at risk of transmitting GBS to their infant and the administration of intrapartum antibiotics have greatly reduced the incidence of neonatal GBS disease. However, there is a requirement for a rapid diagnostic test for GBS that can be carried out in a labour ward setting especially for women whose GBS colonisation status is unknown at the time of delivery. We report the design and evaluation of a real-time PCR test (RiboSEQ GBS test) for the identification of GBS in vaginal swabs from pregnant women. Methods: The qualitative real-time PCR RiboSEQ GBS test was designed based on the bacterial ssrA gene and incorporates a competitive internal standard control. The analytical sensitivity of the test was established using crude lysate extracted from serial dilutions of overnight GBS culture using the IDI Lysis kit. Specificity studies were performed using DNA prepared from a panel of GBS strains, related streptococci and other species found in the genital tract environment. The RiboSEQ GBS test was evaluated on 159 vaginal swabs from pregnant women and compared with the GeneOhm (TM) StrepB Assay and culture for the identification of GBS. Results: The RiboSEQ GBS test is specific and has an analytical sensitivity of 1-10 cell equivalents. The RiboSEQ GBS test was 96.4% sensitive and 95.8% specific compared to &amp;quot;gold standard&amp;quot; culture for the identification of GBS in vaginal swabs from pregnant women. In this study, the RiboSEQ GBS test performed slightly better than the commercial BD GeneOhm (TM) StrepB Assay which gave a sensitivity of 94.6% and a specificity of 89.6% compared to culture. Conclusion: The RiboSEQ GBS test is a valuable method for the rapid, sensitive and specific detection of GBS in pregnant women. This study also validates the ssrA gene as a suitable and versatile target for nucleic acid-based diagnostic tests for bacterial pathogens

Incidence of symptom-driven Coronary Angiographic procedures post-drug-eluting Balloon treatment of Coronary Artery drug-eluting stent in-stent Restenosis-does it matter?

Objectives: The clinical impact of drug-eluting balloon (DEB) coronary intervention for drug-eluting in-stent restenosis (DES-ISR) is not fully known. To further evaluate this impact, we aimed to describe the incidence of symptom-driven coronary angiography (SDCA), an under-reported but potentially informative outcome metric in this cohort of patients. Methods: We retrospectively identified all patients (n=28) who had DEB-treated DES-ISR at University Hospital Limerick in between 2013-2015 and evaluated the incidence of subsequent SDCA as the primary endpoint. Data were expressed as mean &plusmn; SD and %. Results: Baseline demographics demonstrate a mean age 63&plusmn;9 years with 61% of DEB-treated DES-ISR presenting with acute coronary syndrome. Mean number of ISR per patient and number of DEB per lesion was 1.2&plusmn;0.6 lesions and 1.2&plusmn;0.6 balloons, respectively. The incidence of SDCA was 54% after mean follow-up duration of 179&plusmn;241 days. 67.8% of patients had follow-up data beyond 12 months. Within the first year of follow-up, the incidence of SDCA with and without target lesion revascularization (TLR) was 11% and 36% respectively. Among patients with SDCA without TLR, 30% had an acute coronary syndrome not requiring percutaneous coronary intervention. Conclusions: A high incidence of SDCA was observed, particularly within the first 12 months after DEB-treated DES-ISR. This under-reported metric may represent a cohort at higher cardiovascular risk but requires further confirmation in larger studies

MELLO: Study Protocol: A Feasibility Randomised Controlled Trial Of Listening Based Leisure Activities Following Ischemic Stroke

Background and aims: Stroke is the leading cause of disability in older adults. Early post-stroke interventions often focus on physical recovery. Less attention is paid to mood and cognition enhancing interventions that have the potential to improve well-being without the adverse side effects associated with pharmacological interventions. Daily music listening is an accessible and a low cost activity that has been suggested to have a beneficial effect on cognition and mood post stroke. The mechanism of this effect, or if it is reliable, however is not clear. It is speculated that music listening may enhance control of attention in a similar way to mindfulness interventions, that have been demonstrated to be beneficial in the treatment of mood disorders. This study aims to investigate the feasibility and acceptability of a novel intervention combining music listening with brief mindfulness training, within a randomised controlled trial (RCT) context, early post stroke. Method: This is a three-arm, parallel group, single-blind pilot randomised controlled trial (RCT). Individuals with ischemic stroke undergo assessment of mood and cognition (attention and memory) within four weeks post-stroke prior to being randomised to receive an 8-week music listening, mindful music listening, or audiobook listening intervention. Follow-up assessments of mood and cognition are carried out at 3-months and 6-months post-stroke. In addition, a qualitative interview exploring participants’ experience will be completed post intervention. Conclusions: If the mindful music listening intervention is found to be feasible and acceptable, a full scale RCT to investigate its efficacy would be warranted

Inward rectifier potassium conductance regulates membrane potential of canine colonic smooth muscle

The membrane potential of gastrointestinal smooth muscles determines the open probability of ion channels involved in rhythmic electrical activity. The role of Ba2+-sensitive K+ conductances in the maintenance of membrane potential was examined in canine proximal colon circular muscle.Application of Ba2+ (1-100 μM) to strips of tunica muscularis produced depolarization of cells along the submucosal surface of the circular muscle layer. Significantly higher concentrations of Ba2+ were needed to depolarize preparations from which the submucosal and myenteric pacemaker regions were removed.Elevation of extracellular [K+]o (from 5·9 to 12 mM) brought membrane potentials closer to EK (the Nernst potential for K+ ions), suggesting activation of a K+ conductance. This occurred at potentials much more negative than the activation range for delayed rectifier channels (Kv).Forskolin (1 μM) caused hyperpolarization and a leftward shift in the dose-response relationship for Ba2+, suggesting that forskolin may activate a Ba2+-sensitive conductance.Patch-clamp recordings from interstitial cells of Cajal (ICC) revealed the presence of a Ba2+-sensitive inward rectifier potassium conductance. Far less of this conductance was present in smooth muscle cells.Kir2.1 was expressed in the circular muscle layer of the canine proximal colon, duodenum, jejunum and ileum. Kir2.1 mRNA was expressed in greater abundance along the submucosal surface of the circular muscle layer in the colon.These results demonstrate that ICC express a Ba2+-sensitive conductance (possibly encoded by Kir2.1). This conductance contributes to the generation and maintenance of negative membrane potentials between slow waves

A comparison of standard and novel bronchoscopic endobronchial biopsy retrieval methods

Background: The targets of bronchoscopic biopsy now include not only adequate tissue for histologic diagnosis but also tissue for further analysis. We prospectively compared standard and novel bronchoscopic endobronchial biopsy (EBB) retrieval methods attempting to increase tissue yield. Methods: EBB samples were retrieved using techniques A, B, and C using a standard forceps. Method A is routinely performed conventional method, where as in method B, biopsy forceps was left protruded from the bronchoscope and in method C, both valve and forceps were removed to prevent the loss of specimen. At least 6 EBB were retrieved per patient. Results were compared with gold standard composite of confirmatory pathological or clinic-radiologic follow up. Results: A total of 42 of 43 patients completed the study. The final gold standard diagnosis was cancer [non-small cell lung cancer, metastatic, carcinoid, carcinoma in situ (24)], benign disease [sarcoid, amyloid, hamartoma, and chondroid tumor (4)], and benign/nonspecific inflammation (14). EBB retrieved using standard method A were smaller than novel methods B and C (P=0.03). However, the percentage of cases where blood was the predominant component ( > 50%) was less by standard methods A (4/42) than B (16/42) and C (20/42) (P=0.001). There was no difference in mean viable tumor area (n=23, sensitivity for EBB for cancer 96%) between groups A compared with B and C (P 0.27) and adequacy in benign cases by subepithelial depth ( > 0.3 mm) (P=0.38). Conclusion: Standard retrieval of endobronchial biopsies through the bronchoscope and cap does not reduce the size of viable tissue and reduces contaminating blood and necrotic material

MELLO: Study Protocol: A Feasibility Randomised Controlled Trial Of Listening Based Leisure Activities Following Ischemic Stroke

Background and aims: Stroke is the leading cause of disability in older adults. Early post-stroke interventions often focus on physical recovery. Less attention is paid to mood and cognition enhancing interventions that have the potential to improve well-being without the adverse side effects associated with pharmacological interventions. Daily music listening is an accessible and a low cost activity that has been suggested to have a beneficial effect on cognition and mood post stroke. The mechanism of this effect, or if it is reliable, however is not clear. It is speculated that music listening may enhance control of attention in a similar way to mindfulness interventions, that have been demonstrated to be beneficial in the treatment of mood disorders. This study aims to investigate the feasibility and acceptability of a novel intervention combining music listening with brief mindfulness training, within a randomised controlled trial (RCT) context, early post stroke. Method: This is a three-arm, parallel group, single-blind pilot randomised controlled trial (RCT). Individuals with ischemic stroke undergo assessment of mood and cognition (attention and memory) within four weeks post-stroke prior to being randomised to receive an 8-week music listening, mindful music listening, or audiobook listening intervention. Follow-up assessments of mood and cognition are carried out at 3-months and 6-months post-stroke. In addition, a qualitative interview exploring participants’ experience will be completed post intervention. Conclusions: If the mindful music listening intervention is found to be feasible and acceptable, a full scale RCT to investigate its efficacy would be warranted