Rapid and sensitive direct detection and identification of poliovirus from stool and environmental surveillance samples using nanopore sequencing

Global poliovirus surveillance involves virus isolation from stool and environmental samples, intratypic differential (ITD) by PCR, and sequencing of the VP1 region to distinguish vaccine (Sabin), vaccine-derived, and wild-type polioviruses and to ensure an appropriate response. This cell culture algorithm takes 2 to 3 weeks on average between sample receipt and sequencing. Direct detection of viral RNA using PCR allows faster detection but has traditionally faced challenges related to poor sensitivity and difficulties in sequencing common samples containing poliovirus and enterovirus mixtures. We present a nested PCR and nanopore sequencing protocol that allows rapid (99.9%. This novel method shows promise as a faster and safer alternative to cell culture for the detection and real-time sequencing of polioviruses in stool and environmental samples

Effect of early tranexamic acid administration on mortality, hysterectomy, and other morbidities in women with post-partum haemorrhage (WOMAN): an international, randomised, double-blind, placebo-controlled trial

Background Post-partum haemorrhage is the leading cause of maternal death worldwide. Early administration of tranexamic acid reduces deaths due to bleeding in trauma patients. We aimed to assess the effects of early administration of tranexamic acid on death, hysterectomy, and other relevant outcomes in women with post-partum haemorrhage. Methods In this randomised, double-blind, placebo-controlled trial, we recruited women aged 16 years and older with a clinical diagnosis of post-partum haemorrhage after a vaginal birth or caesarean section from 193 hospitals in 21 countries. We randomly assigned women to receive either 1 g intravenous tranexamic acid or matching placebo in addition to usual care. If bleeding continued after 30 min, or stopped and restarted within 24 h of the first dose, a second dose of 1 g of tranexamic acid or placebo could be given. Patients were assigned by selection of a numbered treatment pack from a box containing eight numbered packs that were identical apart from the pack number. Participants, care givers, and those assessing outcomes were masked to allocation. We originally planned to enrol 15 000 women with a composite primary endpoint of death from all-causes or hysterectomy within 42 days of giving birth. However, during the trial it became apparent that the decision to conduct a hysterectomy was often made at the same time as randomisation. Although tranexamic acid could influence the risk of death in these cases, it could not affect the risk of hysterectomy. We therefore increased the sample size from 15 000 to 20 000 women in order to estimate the effect of tranexamic acid on the risk of death from post-partum haemorrhage. All analyses were done on an intention-to-treat basis. This trial is registered with ISRCTN76912190 (Dec 8, 2008); ClinicalTrials.gov, number NCT00872469; and PACTR201007000192283. Findings Between March, 2010, and April, 2016, 20 060 women were enrolled and randomly assigned to receive tranexamic acid (n=10 051) or placebo (n=10 009), of whom 10 036 and 9985, respectively, were included in the analysis. Death due to bleeding was significantly reduced in women given tranexamic acid (155 [1·5%] of 10 036 patients vs 191 [1·9%] of 9985 in the placebo group, risk ratio [RR] 0·81, 95% CI 0·65–1·00; p=0·045), especially in women given treatment within 3 h of giving birth (89 [1·2%] in the tranexamic acid group vs 127 [1·7%] in the placebo group, RR 0·69, 95% CI 0·52–0·91; p=0·008). All other causes of death did not differ significantly by group. Hysterectomy was not reduced with tranexamic acid (358 [3·6%] patients in the tranexamic acid group vs 351 [3·5%] in the placebo group, RR 1·02, 95% CI 0·88–1·07; p=0·84). The composite primary endpoint of death from all causes or hysterectomy was not reduced with tranexamic acid (534 [5·3%] deaths or hysterectomies in the tranexamic acid group vs 546 [5·5%] in the placebo group, RR 0·97, 95% CI 0·87-1·09; p=0·65). Adverse events (including thromboembolic events) did not differ significantly in the tranexamic acid versus placebo group. Interpretation Tranexamic acid reduces death due to bleeding in women with post-partum haemorrhage with no adverse effects. When used as a treatment for postpartum haemorrhage, tranexamic acid should be given as soon as possible after bleeding onset. Funding London School of Hygiene & Tropical Medicine, Pfizer, UK Department of Health, Wellcome Trust, and Bill & Melinda Gates Foundation

Thyroid Dysfunction and Vitamin D Deficiency among Females of Punjab, Pakistan; A Cross Sectional Analysis

Background: Thyroid gland is an important gland which plays a vital role in the stimulation of normal growth and central nervous system (CNS), metabolism regulation, elevated vitamin requirements, metabolism of phosphorus and calcium, promote sexual metabolism, increases mitochondrial metabolism, stimulates the adrenergic activity with myocardial contractility and increase heart rate. In this era, one of the important health issues is vitamin D and calcium deficiency. A large number of populations in all over world are vitamin D or calcium deficient or insufficient. The objective of this study was to evaluate the prevalence of thyroid dysfunction and vitamin D deficiency in females.Methods: A cross sectional study was designed to check the prevalence of thyroid disease and its correlation with vitamin D levels in females. Venous blood was drawn from the female’s patients (11—80 years of age) using gel disposable vials (3.5 ml) in aseptic condition. Samples were centrifuged at four thousand revolutions per minute for five minutes and serums were separated. After the separation of serum, the samples were transferred to the laboratory for the automated estimation of Thyroid Stimulating Hormone (TSH), Triiodothyronine (T3), Thyroxine and Vitamin D.Results: It was observed that out of 79 females who had gone through thyroid profile testing, 70% females had normal thyroid profile. However, hypothyroidism was found in 23% females and 7% females had the condition of hyperthyroidism. Out of 18 (23%) reported cases of hypothyroidism, 8 cases were of mild (subclinical) hypothyroidism, 3 cases indicated non-thyroidal illness; rare pituitary hypothyroidism. Out of 79 female participants, only 20 (25.31%) females had normal serum vitamin D levels. Overall, 59 (74.68%) females had vitamin D deficiency. When the vitamin D deficiency was correlated with thyroid dysfunction, it was observed that vitamin D levels were non-significantly (p = 0.35) associated with hypothyroidism.Conclusion: Hypothyroidism was found prevalent in the tested female population, as 23% of the tested population had hypothyroidism while 9% of them have hyperthyroidism. Moreover, majority of the population had vitamin D deficiency.Keywords: Thyroid Dysfunction; Vitamin D Deficiency; Females

Evaluation of the bag-mediated filtration system as a novel tool for poliovirus environmental surveillance: Results from a comparative field study in Pakistan

<div><p>Poliovirus (PV) environmental surveillance (ES) plays an important role in the global eradication program and is crucial for monitoring silent PV circulation especially as clinical cases decrease. This study compared ES results using the novel bag-mediated filtration system (BMFS) with the current two-phase separation method. From February to November 2016, BMFS and two-phase samples were collected concurrently from twelve sites in Pakistan (<i>n</i> = 117). Detection was higher in BMFS than two-phase samples for each Sabin-like (SL) PV serotype (<i>p</i><0.001) and wild PV type 1 (WPV1) (<i>p</i> = 0.065). Seventeen sampling events were positive for WPV1, with eight discordant in favor of BMFS and two in favor of two-phase. A vaccine-derived PV type 2 was detected in one BMFS sample but not the matched two-phase. After the removal of SL PV type 2 (SL2) from the oral polio vaccine in April 2016, BMFS samples detected SL2 more frequently than two-phase (<i>p</i> = 0.016), with the last detection by either method occurring June 12, 2016. More frequent PV detection in BMFS compared to two-phase samples is likely due to the greater effective volume assayed (1620 mL vs. 150 mL). This study demonstrated that the BMFS achieves enhanced ES for all PV serotypes in an endemic country.</p></div

Detection of wild poliovirus type 1 (WPV1) and vaccine-derived poliovirus type 2 (VDPV2) in bag-mediated filtration system (BMFS) and two-phase samples from 12 study sites in 10 Pakistani districts from February-November 2016.

<p>The genetic cluster of WPV1 isolates is shown. The VDPV2 detected was an ambiguous VDPV2 (aVDPV2) with 12 nucleotide changes.</p

Comparison of PV detection in matching BMFS and two-phase samples as measured by ITD after virus amplification on L20B and RD cells.

<p>Comparison of PV detection in matching BMFS and two-phase samples as measured by ITD after virus amplification on L20B and RD cells.</p

SL2 detection in BMFS and two-phase samples as measured by ITD after virus amplification on L20B and RD cells.

<p>SL2 detection in BMFS and two-phase samples as measured by ITD after virus amplification on L20B and RD cells.</p

Location and type of sampling sites in Pakistan.

<p>Location and type of sampling sites in Pakistan.</p

Detection of poliovirus (PV) in bag-mediated filtration system (BMFS) and two-phase samples from 12 study sites in 10 Pakistani districts from February-November 2016.

<p>NPEV is non-polio enterovirus. SL1 is Sabin-like poliovirus type 1. SL2 is Sabin-like poliovirus type 2. SL3 is Sabin-like poliovirus type 3. WPV1 is wild poliovirus type 1. VDPV2 is vaccine-derived poliovirus type 2.</p