Polymerase Chain Reaction Screening Research Articles

Transcriptome Analysis Reveals Cross-Talk between the Flagellar Transcriptional Hierarchy and Secretion System in Plesiomonas shigelloides

Plesiomonas shigelloides, a Gram-negative bacillus, is the only member of the Enterobacteriaceae family able to produce polar and lateral flagella and cause gastrointestinal and extraintestinal illnesses in humans. The flagellar transcriptional hierarchy of P. shigelloides is currently unknown. In this study, we identified FlaK, FlaM, FliA, and FliAL as the four regulators responsible for polar and lateral flagellar regulation in P. shigelloides. To determine the flagellar transcription hierarchy of P. shigelloides, the transcriptomes of the WT and ΔflaK, ΔflaM, ΔfliA, and ΔfliAL were carried out for comparison in this study. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and luminescence screening assays were used to validate the RNA-seq results, and the Electrophoretic Mobility Shift Assay (EMSA) results revealed that FlaK can directly bind to the promoters of fliK, fliE, flhA, and cheY, while the FlaM protein can bind directly to the promoters of flgO, flgT, and flgA. Meanwhile, we also observed type VI secretion system (T6SS) and type II secretion system 2 (T2SS-2) genes downregulated in the transcriptome profiles, and the killing assay revealed lower killing abilities for ΔflaK, ΔflaM, ΔfliA, and ΔfliAL compared to the WT, indicating that there was a cross-talk between the flagellar hierarchy system and bacterial secretion system. Invasion assays also showed that ΔflaK, ΔflaM, ΔfliA, and ΔfliAL were less effective in infecting Caco-2 cells than the WT. Additionally, we also found that the loss of flagellar regulators causes the differential expression of some of the physiological metabolic genes of P. shigelloides. Overall, this study aims to reveal the transcriptional hierarchy that controls flagellar gene expression in P. shigelloides, as well as the cross-talk between motility, virulence, and physiological and metabolic activity, laying the groundwork for future research into P. shigelloides’ coordinated survival in the natural environment and the mechanisms that infect the host.

The ion channel TRPV5 regulates B-cell signaling and activation.

B-cell activation triggers the release of endoplasmic reticulum calcium stores through the store-operated calcium entry (SOCE) pathway resulting in calcium influx by calcium release-activated calcium (CRAC) channels on the plasma membrane. B-cell-specific murine knockouts of SOCE do not impact humoral immunity suggesting that alternative channels may be important. We identified a member of the calcium-permeable transient receptor potential (TRP) ion channel family, TRPV5, as a candidate channel expressed in B cells by a quantitative polymerase chain reaction (qPCR) screen. To further investigate the role of TRPV5 in B-cell responses, we generated a murine TRPV5 knockout (KO) by CRISPR-Cas9. We found TRPV5 polarized to B-cell receptor (BCR) clusters upon stimulation in a PI3K-RhoA-dependent manner. TRPV5 KO mice have normal B-cell development and mature B-cell numbers. Surprisingly, calcium influx upon BCR stimulation in primary TRPV5 KO B cells was not impaired; however, differential expression of other calcium-regulating proteins, such as ORAI1, may contribute to a compensatory mechanism for calcium signaling in these cells. We demonstrate that TRPV5 KO B cells have impaired spreading and contraction in response to membrane-bound antigen. Consistent with this, TRPV5 KO B cells have reduced BCR signaling measured through phospho-tyrosine residues. Lastly, we also found that TRPV5 is important for early T-dependent antigen specific responses post-immunization. Thus, our findings identify a role for TRPV5 in BCR signaling and B-cell activation.

Management outcome of a COVID-19 Positive Child with Multiple Contacts admitted in a General Intensive Care Unit-A Case Report

COVID-19 infection in paediatric patients is relatively low in Nigeria. The low index of suspicion made healthcare providers to be at risk of contracting the disease from this age group making prevention and contact tracing difficult. We report the management of a 9-month- old male child who had palatoplasty following a prior negative polymerase chain reaction (PCR) screening. The patient underwent palatoplasty for isolated cleft palate under general anaesthesia and endotracheal intubation. The patient was found to be desaturating significantly on admission to the recovery room and was subsequently taken back to the theatre with multiple attempts at intubation. A repeat PCR done was positive and subsequently, he was admitted into the ICU for ventilatory support. A chest radiograph taken on admission revealed features of severe bronchopneumonia and COVID-19 pneumonic changes. He was ventilated for 14 days and thereafter, was discharged home on a tracheostomy in good clinical condition. The mother of the child and 20 members of staff (23.5%) out of the 85 personnel who had close contact with the child in the course of treatment tested PCR positive. Contact tracing was difficult among those infected. They all recovered from the disease without complications. The treatment outcome in this COVID-19-positive child was good and contact tracing was effective in reducing the transmission of SARS-CoV-2 and limiting the impact of the outbreak. We hereby recommend preventive measures against the infection, vaccination and establishment of dedicated paediatric ICU for infectious diseases.

Investigation of Trypanosoma evansi in Sumatran Elephant (Elephas maximus sumatranus) in Indonesia Using Various Methods

Trypanosoma evansi is a widespread hemoprotozoa that causes trypanosomiasis in both livestock and wildlife. One of the susceptible animals is the Sumatran elephant, which is included in the endangered species category. The infection of this parasite often develops into chronic and sub-clinical forms in elephants, therefore it may become unnoticed and hard to diagnose. This study aimed to analyze and evaluate the infection of T. evansi in semi-captive Sumatran elephants in Way Kambas National Park, Indonesia, using various diagnostic tests. The prevalence of T. evansi in a total of 53 Sumatran elephants was estimated using a card agglutination test for trypanosomiasis (CATT) in 2016. A longitudinal study was later conducted in 2019 using Giemsa stained blood smear (GSBS) and polymerase chain reaction (PCR). The results showed that 26.4% of samples (14/53) were positively detected by both CATT in 2016 and PCR in 2019, while GSBS was unable to detect the parasites in all samples. Furthermore, four individuals were confirmed to have persistent infections. This study concluded that the ability of CATT and PCR were more convincing over GSBS for the diagnosis of sub-clinical trypanosomiasis in Sumatran elephants. However, it is recommended to use a combination of CATT as a screening tool and PCR as a confirmatory test for reliable results.

The Larynx is Protected from Secondary and Vertical Papillomavirus Infection in Immunocompetent Mice.

Mouse papillomavirus MmuPV1 causes both primary and secondary infections of the larynx in immunocompromised mice. Understanding lateral and vertical transmission of papillomavirus to the larynx would benefit patients with recurrent respiratory papillomatosis (RRP). To test the hypothesis that the larynx is uniquely vulnerable to papillomavirus infection, and to further develop a mouse model of RRP, we assessed whether immunocompetent mice were vulnerable to secondary or vertical laryngeal infection with MmuPV1. Larynges were collected from 405 immunocompetent adult mice that were infected with MmuPV1 in the oropharynx, oral cavity, or anus, and 31 mouse pups born to immunocompetent females infected in the cervicovaginal tract. Larynges were analyzed via polymerase chain reaction (PCR) of lavage fluid or whole tissues for viral DNA, histopathology, and/or in situ hybridization for MmuPV1 transcripts. Despite some positive laryngeal lavage PCR screens, all laryngeal tissue PCR and histopathology results were negative for MmuPV1 DNA, transcripts, and disease. There was no evidence for lateral spread of MmuPV1 to the larynges of immunocompetent mice that were infected in the oral cavity, oropharynx, or anus. Pups born to infected mothers were negative for laryngeal MmuPV1 infection from birth through weaning age. Secondary and vertical laryngeal MmuPV1 infections were not found in immunocompetent mice. Further work is necessary to explore immunologic control of laryngeal papillomavirus infection in a mouse model and to improve preclinical models of RRP. NA Laryngoscope, 134:2322-2330, 2024.

A cross-sectional analysis identifies a low prevalence of Plasmodium ovale species infections in symptomatic and asymptomatic individuals in Kilifi county, Kenya.

Background The focus on P. falciparum diagnosis has led to an underestimation of the global burden of malaria resulting from neglected Plasmodium species. However, there is still scarce data on the prevalence of P. ovale globally. To address this knowledge gap, data collected from cross-sectional studies in Kilifi county were used to: 1) determine the prevalence of P. ovale species infections; and 2) determine the sensitivity of different diagnostic assays in detecting P. ovale species infections. Methods A total of 531 individuals were sampled across three study sites in Kilifi County, Kenya between 2009 and 2020. Blood smears were prepared from peripheral blood and screened for Plasmodium parasite stages using light microscopy. Molecular screening involved DNA extraction of dried blood spots and blood in ethylenediaminetetraacetic acid, polymerase chain reaction (PCR) using primers targeting the 18 small ribosomal subunit and sequencing. Results Microscopy screening revealed that the most prevalent species was P. falciparum (32.0%) followed by P. malariae (9.0%) and then P. ovale (1.5%). PCR screening identified additional P. ovale species positives cases. Overall PCR results indicate that43 (8.1%) out of the 531 individuals harbored P. ovale curtisi infection with the highest prevalence reported in the tertiary health facility, (14.6%, 95% CI 8-23.6%), followed by the primary health facility (8.3%, 95% CI 5.4-11.9%), and the community from a cross-sectional blood survey, (3.6%, 95% CI 1.2-8.2%). Microscopy screening for P. ovale had a low sensitivity of 7% (95% CI 1-19-30%) and a high specificity of 99% (95% CI 98-100%). Sequencing results confirmed the presence of P.ovale curtisi. Conclusions This study provides baseline data for P.ovale species surveillance in Kilifi County, primarily using PCR to improve diagnosis. These results suggest that malaria elimination and eradication efforts should not only concentrate on P. falciparum but should embrace a holistic approach towards elimination of all Plasmodium species.

SARS-CoV-2 RT-PCR as a universal screening on planned admission in asymptomatic patients

Universal screening for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on admission is reportedly beneficial in preventing nosocomial infections. However, some issues remain, including low positivity rate, cost, and time required for testing. We describe SARS-CoV-2 reverse transcription polymerase chain reaction (PCR) for universal screening in asymptomatic patients on planned admissions. In total, 14,574 patients were included between October 12, 2020, and June 23, 2022. The PCR-positive rate for the period was 0.44 % (64/14,574). The PCR positivity for the epidemic period by strain was 0.28 % (95 % confidence interval [CI] 0.12–0.56 %), 0.16 % (95 % CI 0.05–0.37 %), 0.21 % (95 % CI 0.09–0.41 %), and 0.9 % (95 % CI 0.65–1.2 %) for the wild-type strain, Alpha, Delta, and Omicron variants, respectively. The proportion of Ct values < 30 was higher in the first half of the epidemic (first vs. second, 29.4 % [95 % CI 16.9–44.8 %] vs. 16.7 % [95 % CI 6.0–28.5 %]), whereas that of Ct values ≥ 35 increased significantly in the second half (first vs. second, 32.4 % [95 % CI 19.3–47.8 %] vs. 70.0 % [95 % CI 53.5–83.4 %]). Of all positives, 50 % (32/64) had a coronavirus disease (COVID-19) history before PCR screening, with a median of 28 days (10–105) from COVID-19 onset or positive to PCR screening. PCR screening may help detect positives with high viral loads early in the epidemic for each mutant strain, with an increasing proportion of positives with low viral loads later in the epidemic. PCR testing may be unnecessary for recently diagnosed cases and patients in whom reinfection is unlikely.

2073. Impact of in-house Candida auris polymerase chain reaction (PCR) screening on admission on the incidence rates of surveillance and clinical cultures with C. auris and associated cost-savings

Abstract Background Candida auris (CA) transmission has significantly increased in the United States. Routine screening by PCR methods and economic impact have not been previously defined. We present our experience implementing in-house PCR testing on admission for screening of colonization with CA in an area of high prevalence. Methods Study conducted across the acute care and inpatient rehabilitation hospitals at a large integrated, university affiliated, public health system in Miami, Florida. The pre-intervention (August 1, 2019 -July 31, 2021) testing was sent out to a refence laboratory, and post-intervention (August 1, 2021 - January 31, 2023) testing was performed in-house. Our screening, testing and isolation protocols are presented in Figure 1A and 1B. Interrupted time series was done to analyze changes in the incidence rates of CA present on admission (CA-POA) and hospital-onset fungemia (CA-HOF), and the estimated economic impact on isolation costs and testing were calculated. Candida auris screening and isolation process Figures describing the 2-step screening process for Candida auris, including the questionnaire on the electronic medical record, and the transmission-based precautions followed Results A total of 4,478 PCRs were done on 4,270 patients. We identified 159 unique CA-POA. During the pre-intervention period there was a baseline trend towards a monthly increase in the incidence rate of CA-POA of 3% (95% CI 1.00-1.06; p=0.11). Following the intervention there was an immediate change with an incidence rate ratio of 2.57 (95% CI 1.16-5.69, p=0.02) compared to the pre-intervention period (Figure 2A). Seventy-five unique cases of CA-HOF were identified. In the pre-intervention period, the baseline rate of CA-HOF was increasing monthly by 14% (95% CI 1.05-1.24; p=0.002). No immediate change in rates was seen after introducing in-house testing; throughout the post-intervention period there was a change in slope showing a monthly decrease in rates of 13% (95% CI 0.80-0.99; p=0.02) (Figure 2B). A preliminary economic impact analysis is presented in Table 1. Interrupted Time Series Analysis of rates of Candida auris present on admission and Candida auris hospital-onset fungemia Impact of performing screening for C. auris present on admission (POA) using in-house PCR testing had on incidence rates of: A) C. auris present on admission per 10,000 patient-admissions, and, B) C. auris hospital-onset fungemia per 10,000 patient-days. Circles represent observed rates. Solid lines estimated from the interrupted time series model; segmented line represent the counterfactual model without the effect of the intervention. Table summarizing the costs and savings associated with the intervention Conclusion By performing in-house PCR colonization screening on admission for the detection of CA, we found a doubling of CA-POA rates, and a subsequent decrease in the incidence rates of CA-HOF. In-house testing was cost effective in our setting, with savings estimates ranging from $21,441.40 to $95,240.08, and should be considered in areas of high prevalence to decrease invasive infections and prevent horizontal transmission. Disclosures Lilian M. Abbo, MD, MBA, Ferring: Advisor/Consultant|Pfizer: Advisor/Consultant|Regeneron: Grant/Research Support|Shionogi: Advisor/Consultant

Impact of In-house Candida auris Polymerase Chain Reaction Screening on Admission on the Incidence Rates of Surveillance and Blood Cultures With C. auris and Associated Cost Savings.

The impact of strategies for rapid diagnostic screening of Candida auris on hospital operations has not been previously characterized. We describe the implementation of in-house polymerase chain reaction (PCR) testing on admission for screening of colonization with C. auris, associated process improvements, and financial impact. This study was conducted across an integrated health system. Patients were tested based on risk factors for C. auris carriage. Pre-intervention, the PCR was sent out to a reference laboratory, and postintervention was performed in-house. Changes in the incidence rates (IRs) of C. auris present on admission (CA-POA) and C. auris hospital-onset fungemia (CA-HOF) were assessed using interrupted time series analysis. The economic impact on isolation and testing costs was calculated. Postintervention, the IR of CA-POA doubled (IRR, 2.57; 95% CI, 1.16-5.69; P = .02) compared with the pre-intervention period. The baseline rate of CA-HOF was increasing monthly by 14% (95% CI, 1.05-1.24; P = .002) pre-intervention, while during the postintervention period there was a change in slope with a monthly decrease in IR of 13% (95% CI, 0.80-0.99; P = .02). The median turnaround time (TAT) of the results (interquartile range) was reduced from 11 (8-14) days to 2 (1-3) days. Savings were estimated to be between $772 513.10 and $3 730 480.26. By performing in-house PCR for screening of C. auris colonization on admission, we found a doubling of CA-POA rates, a subsequent decrease in CA-HOF rates, reduced TAT for PCR results, and more efficient use of infection control measures. In-house testing was cost-effective in a setting of relatively high prevalence among individuals with known risk factors.

Prevalence and Antimicrobial Resistance of Salmonella Dublin and Thermotolerant Campylobacter in Liver from Veal Calves in Québec, Canada.

Salmonella Dublin and Campylobacter spp. are two foodborne pathogens of importance. A small number of studies reported that consumption of veal liver was associated with an increased risk of human illness from these two pathogens. To better characterize the risk of exposure from liver, a cross-sectional study was conducted to estimate the prevalence of white veal calf liver contamination with these two pathogens and to characterize the antimicrobial non-susceptibility patterns of isolates. Veal liver samples were collected at two slaughterhouses in Quebec, Canada, in 2016 and 2017. Samples were submitted for polymerase chain reaction (PCR) screening followed by culture of Salmonella and thermotolerant Campylobacter. Isolates were tested for antimicrobial susceptibility using broth microdilution. Salmonella Dublin was the only serotype cultured from 3.6% (95% confidence interval [CI]: 0.0-7.9) of 560 liver samples. Among them and for technical reasons, 498 were tested by PCR for Campylobacter. The prevalence of PCR-positive livers was estimated to be 65.8% (95% CI: 58.7-72.9) for Campylobacter jejuni and 7.0% (95% CI: 3.9-10.1%) for Campylobacter coli. Fourteen Salmonella Dublin isolates were submitted for antimicrobial resistance (AMR) testing; all were non-susceptible to at least eight antimicrobials from six different classes. Most (81.4%) of the 188 C. jejuni isolates submitted for AMR testing were non-susceptible to tetracycline, and 23.0% of isolates were non-susceptible to nalidixic acid and ciprofloxacin. Of the seven C. coli isolates, four were multidrug resistant. This study highlights the importance of veal liver as a potential source of exposure to multidrug-resistant Salmonella Dublin and thermotolerant Campylobacter spp.

A cross-sectional analysis identifies a low prevalence of Plasmodium ovale species infections in symptomatic and asymptomatic individuals in Kilifi county, Kenya.

Background: The focus on P. falciparum diagnosis has led to an underestimation of the global burden of malaria resulting from neglected Plasmodium species. However, there is still scarce data on the prevalence of P. ovale globally. To address this knowledge gap, data collected from cross-sectional studies in Kilifi county were used to: 1) determine the prevalence of P. ovale species infections; and 2) determine the sensitivity of different diagnostic assays in detecting P. ovale species infections. Methods: A total of 531 individuals were sampled across three study sites in Kilifi County, Kenya between 2009 and 2020. Blood smears were prepared from peripheral blood and screened for Plasmodium parasite stages using light microscopy. Molecular screening involved DNA extraction of dried blood spots and blood in ethylenediaminetetraacetic acid, polymerase chain reaction (PCR) using primers targeting the 18 small ribosomal subunit and sequencing. Results: Microscopy screening revealed that the most prevalent species was P. falciparum (32.0%) followed by P. malariae (9.0%) and then P. ovale (1.5%). PCR screening identified additional P. ovale species positives cases. Overall, 48 (8.2%) out of the 531 individuals harbored P. ovale species infection with the highest prevalence reported in the tertiary health facility, (14.6%, 95% CI 8-23.6%), followed by the primary health facility (8.6%, 95% CI 5.4-11.9%), and the community from a cross-sectional blood survey, (6.5%, 95% CI 3.0-11.8%). Microscopy screening for P. ovale had a low sensitivity of 7% (95% CI 1-19-30%) and a high specificity of 99% (95% CI 98-100%). Sequencing results confirmed the presence of P.ovale curtisi. Conclusions: This study provides baseline data for P. ovale species surveillance in Kilifi County, primarily using PCR to improve diagnosis. These results suggest that malaria elimination and eradication efforts should not only concentrate on P. falciparum but should embrace a holistic approach towards elimination of all Plasmodium species.

Molecular detection and viability discrimination of zoonotic protozoan pathogens in oysters and seawater

The presence of foodborne protozoan pathogens including Cryptosporidium parvum, Giardia duodenalis, Toxoplasma gondii, and Cyclospora cayetanensis in commercial shellfish has been reported across diverse geographical regions. In the present study, a novel multiplex nested polymerase chain reaction (PCR) assay was validated to simultaneously detect and discriminate these four targeted parasites in oyster tissues including whole tissue homogenate, digestive gland, gills, and hemolymph, as well as seawater where shellfish grow. To differentiate viable and non-viable protozoan (oo)cysts, we further evaluated reverse transcription quantitative PCR (RT-qPCR) assays through systematic laboratory spiking experiments by spiking not only dilutions of viable parasites but also mixtures of viable and non-viable parasites in the oyster tissues and seawater. Results demonstrate that multiplex PCR can detect as few as 5–10 (oo)cysts in at least one oyster matrix, as well as in 10 L of seawater. All parasites were detected at the lowest spiking dilution (5 (oo)cysts per extract) in hemolymph, however the probability of detection varied across the difference matrices tested for each parasite. RT-qPCR further discriminated viable from non-viable (heat-inactivated) C. parvum and T. gondii in seawater and hemolymph but did not perform well in other oyster matrices. This systematic spiking study demonstrates that a molecular approach combining multiplex PCR for sensitive and affordable screening of protozoan DNA and subsequent RT-qPCR assay for viability discrimination presents an important advance for accurately determining the risk of protozoal illness in humans due to consumption of contaminated shellfish.

Using Methicillin-Resistant Staphylococcus aureus Nasal Screens to Rule Out Methicillin-Resistant S aureus Pneumonia in Surgical Intensive Care Units

IntroductionThe methicillin-resistant Staphylococcus aureus (MRSA) polymerase chain reaction (PCR) has a high negative predictive value (NPV). We aimed to understand if there was a difference in the NPV of the MRSA screen in surgical intensive care units (ICUs) and to determine its role in antibiotic de-escalation. MethodsWe performed a single-center, retrospective cohort study of adults with a positive respiratory culture and MRSA nasal PCR admitted to a surgical ICU from 2016 to 2019. Patients were stratified by surgical ICU: cardiothoracic/cardiovascular intensive care unit (CVICU) or transplant/acute care surgery intensive care unit (ACS-ICU). Our primary outcome was the NPV of MRSA screen. Secondary outcome was the duration of empiric MRSA-targeted therapy. ResultsWe analyzed 61 patients: 42.6% (n = 26) ACS-ICU and 57.4% (n = 35) CVICU. There were no differences in age, comorbidities, prior MRSA infection, recent antibiotic use, immunocompromised status, or renal replacement therapy. At pneumonia diagnosis, more patients in the ACS-ICU were hospitalized ≥5 d (65.4% versus 8.6%, P < 0.0001) and more patients in the CVICU were in septic shock (88.6% versus 34.5%, P < 0.0001) and thrombocytopenic (40% versus 11.5%, P = 0.02). NPV of the PCR was similar (ACS-ICU: 0.92 [0.75-0.98], CV-ICU 0.89 [0.73-0.96]). On multivariable linear regression, the CVICU was associated with longer empiric therapy (β 1.5, 95% CI 0.8-2.3, P < 0.0001), as was hospitalization for ≥5 d (β 0.73, 95% CI 0.06-1.39, P = 0.03). ConclusionsThe MRSA nasal PCR screen has a high NPV for ruling out MRSA pneumonia in critically ill surgical patients. However, patients in the CVICU and patients hospitalized ≥5 d had a longer time to de-escalation of MRSA-targeted therapy, potentially due to higher clinical risk profile.

REVERSIBLE LUNG FIBROSIS FOLLOWING SEVERE COVID-19 INFECTIONA CASE REPORT

After wide spread of COVID-19 pneumonia with thousands of cases complicated with acute respiratory distress syndrome (ARDS) of which high percentage fully recovered and resumed their normal life without any disability. It was known that those patients will have residual fibrosis which may affect their daily activity on the long run.There was always paramount question about the reversibility of such fibrosis especially with well-known concept about absence of any radical treatment for such etiology.We present a case of 54 years old male patient, non-smoker with irrelevant medical history apart from well controlled hypertension. He was referred to our facility with fever, productive cough, and shortness of breath. An initial polymerase chain reaction (PCR) screening for SARS-COV2 was positive. A high-resolution CT (HRCT) chest revealed Multi-focal patchy air-space ground glass density with peripheral and basal predominance more affecting the left side, suggestive of viral pneumonia. The patient was commenced on low flow oxygen therapy and symptomatic treatment. Four days later the symptoms aggravated and oxygen requirement increased to non-rebreathing oxygen mask (NRM). We treated him according to COVID protocol withtherapeutic anticoagulation, corticosteroids and oxygen therapy which was adjusted to maintain saturation 92%, but his condition deteriorated and was shifted to intensive care unit. Later patient improved andwas shifted back to the medical ward. We were able to wean him off oxygen and he was discharged home oncorticosteroidsand scheduled for a follow up visit in the pulmonology clinic. Repeated HRCT chest showed regressive course concerning both the airway space disease and the fibrotic changes. The patient returned to his normal life activity with no residual limitation of his physical activity or development of shortness of breath with exertion. Latest HRCT was completely normal.

Cost-Effectiveness of Universal Asymptomatic Preoperative SARS-CoV-2 Polymerase Chain Reaction Screening: A Cost-Utility Analysis.

An early report has shown the clinical benefit of the asymptomatic preoperative severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) screening test, and some clinical guidelines recommended this test. However, the cost-effectiveness of asymptomatic screening was not evaluated. We aimed to investigate the cost-effectiveness of universal preoperative screening of asymptomatic patients for SARS-CoV-2 using polymerase chain reaction (PCR) testing. We evaluated the cost-effectiveness of asymptomatic screening using a decision tree model from a payer perspective, assuming that the test-positive rate was 0.07% and the screening cost was 8500 Japanese yen (JPY) (approximately 7601 US dollars [USD]). The input parameter was derived from the available evidence reported in the literature. A willingness-to-pay threshold was set at 5 000 000 JPY/quality-adjusted life-year (QALY). The incremental cost of 1 death averted was 74 469 236 JPY (approximately 566 048 USD) and 291 123 368 JPY/QALY (approximately 2 212 856 USD/QALY), which was above the 5 000 000 JPY/QALY willingness-to-pay threshold. The incremental cost-effectiveness ratio fell below 5 000 000 JPY/QALY only when the test-positive rate exceeded 0.739%. However, when the probability of developing a postoperative pulmonary complication among SARS-CoV-2-positive patients was below 0.22, asymptomatic screening was never cost-effective, regardless of how high the test-positive rate became. Asymptomatic preoperative universal SARS-CoV-2 PCR screening is not cost-effective in the base case analysis. The cost-effectiveness mainly depends on the test-positive rate, the frequency of postoperative pulmonary complications, and the screening costs; however, no matter how high the test-positive rate, the cost-effectiveness is poor if the probability of developing postoperative pulmonary complications among patients positive for SARS-CoV-2 is sufficiently reduced.

Effectiveness and Costs of Molecular Screening and Treatment for Bacterial Vaginosis to Prevent Preterm Birth

Bacterial vaginosis (BV) is a well-known risk factor for preterm birth. Molecular diagnosis of BV is now available. Its impact in the screening and treatment of BV during pregnancy on preterm births has not been evaluated to date. To evaluate the clinical and economic effects of point-of-care quantitative real-time polymerase chain reaction screen and treat for BV in low-risk pregnant women on preterm birth. The AuTop trial was a prospective, multicenter, parallel, individually randomized, open-label, superiority trial conducted in 19 French perinatal centers between March 9, 2015, and December 18, 2017. Low-risk pregnant women before 20 weeks' gestation without previous preterm births or late miscarriages were enrolled. Data were analyzed from October 2021 to November 2022. Participants were randomized 1:1 to BV screen and treat using self-collected vaginal swabs (n = 3333) or usual care (n = 3338). BV was defined as Atopobium vaginae (Fannyhessea vaginae) load of 108 copies/mL or greater and/or Gardnerella vaginalis load of 109 copies/mL or greater, using point-of-care quantitative real-time polymerase chain reaction assays. The control group received usual care with no screening of BV. Overall rate of preterm birth before 37 weeks' gestation and total costs were calculated in both groups. Secondary outcomes were related to treatment success as well as maternal and neonate health. Post hoc subgroup analyses were conducted. Among 6671 randomized women (mean [SD] age, 30.6 [5.0] years; mean [SD] gestational age, 15.5 [2.8] weeks), the intention-to-treat analysis of the primary clinical and economic outcomes showed no evidence of a reduction in the rate of preterm birth and total costs with the screen and treat strategy compared with usual care. The rate of preterm birth was 3.8% (127 of 3333) in the screen and treat group and 4.6% (153 of 3338) in the control group (risk ratio [RR], 0.83; 95% CI, 0.66-1.05; P = .12). On average, the cost of the intervention was €203.6 (US $218.0) per participant, and the total average cost was €3344.3 (US $3580.5) in the screen and treat group vs €3272.9 (US $3504.1) in the control group, with no significant differences being observed. In the subgroup of nulliparous women (n = 3438), screen and treat was significantly more effective than usual care (RR, 0.62; 95% CI, 0.45-0.84; P for interaction = .003), whereas no statistical difference was found in multiparous (RR, 1.30; 95% CI, 0.90-1.87). In this clinical trial of pregnant women at low risk of preterm birth, molecular screening and treatment for BV based on A vaginae (F vaginae) and/or G vaginalis quantification did not significantly reduce preterm birth rates. Post hoc analysis suggests a benefit of screen and treat in low-risk nulliparous women, warranting further evaluation in this group. ClinicalTrials.gov Identifier: NCT02288832.

Performance of Methicillin-Resistant Staphylococcus aureus Polymerase Chain Reaction Nasal Screening for Ruling Out MRSA Pneumonia in Hospitalized, Immunocompromised Patients.

Background: Recent literature demonstrates support for using methicillin-resistant Staphylococcus aureus (MRSA) nasal swab polymerase chain reaction (NaPCR) screening as an antimicrobial stewardship tool aiding early de-escalation of anti-MRSA antimicrobials. However, immunocompromised patients have been underrepresented in previous studies despite increased risk of morbidity and mortality from multidrug-resistant organisms (MDRO). Objective: The purpose of this study was to determine the negative predictive value (NPV) of the MRSA NaPCR in hospitalized, immunocompromised adult patients with suspected pneumonia. Methods: A single-center, retrospective, observational review was conducted of hospitalized, immunocompromised adult patients that had an MRSA NaPCR obtained between March 1, 2020 and January 10, 2021. For inclusion, bacterial cultures must have been collected within 2 weeks after MRSA NaPCR. The primary outcome was the NPV of MRSA NaPCR in hospitalized, immunocompromised patients with suspected pneumonia. Secondary outcomes include NPV in other infections. Results: Between March 1, 2020 and January 10, 2021, 59 patients with 78 unique cultures, including 28 respiratory cultures, were included in the study. The NPV of the MRSA NaPCR for pneumonia was 91.7%. The NPV for bloodstream infections was 100% and for urinary tract infections was 100%, but interpretation of these results should be cautioned due to the small sample sizes. Conclusion: The NPV of MRSA NaPCR in pneumonia remains high in this study. The MRSA NaPCR has utility as a de-escalation tool in hospitalized, immunocompromised adult patients, but larger studies are warranted to evaluate all immunocompromised patient populations.

Tracheal bifurcation repair for blunt thoracic trauma in a patient with COVID-19

BackgroundTracheobronchial injury (TBI) is a rare but potentially life-threatening trauma that requires prompt diagnosis and treatment. We present a case in which a patient with COVID-19 infection was successfully treated for a TBI through surgical repair and intensive care with extracorporeal membrane oxygenation (ECMO) support.Case presentationThis is the case of a 31-year-old man transported to a peripheral hospital following a car crash. Tracheal intubation was performed for severe hypoxia and subcutaneous emphysema. Chest computed tomography showed bilateral lung contusion, hemopneumothorax, and penetration of the endotracheal tube beyond the tracheal bifurcation. A TBI was suspected; moreover, his COVID-19 polymerase chain reaction screening test was positive. Requiring emergency surgery, the patient was transferred to a private negative pressure room in our intensive care unit. Due to persistent hypoxia and in preparation for repair, the patient was started on veno-venous ECMO. With ECMO support, tracheobronchial injury repair was performed without intraoperative ventilation. In accordance with the surgery manual for COVID-19 patients in our hospital, all medical staff who treated this patient used personal protective equipment. Partial transection of the tracheal bifurcation membranous wall was detected and repaired using 4-0 monofilament absorbable sutures. The patient was discharged on the 29th postoperative day without postoperative complications.ConclusionsECMO support for traumatic TBI in this patient with COVID-19 reduced mortality risk while preventing aerosol exposure to the virus.

Current Issues on Zika Virus Disease: The Nigeria Perspective

Zika virus sickness causes a mild fever, rash, headache, arthralgia, myalgia, asthenia, and non-purulent conjunctivitis, and symptoms appear two to seven days after contact with an infected mosquito. The zika virus is what causes the zika virus sickness, or just zika. Seldom do symptoms appear, and when they do, they could be comparable to dengue fever symptoms. Possible symptoms include maculopapular rash, fever, red eyes, joint pain, headaches, and a likely duration of about 2 days. Possible adverse effects include rashes, fever, conjunctivitis, aching muscles and joints, general malaise, and headaches. Zika virus infection has also been strongly linked to Guillain-Barré syndrome (GBS), which is characterised by rapidly developing muscle weakness brought on by immune system injury to the peripheral nerve system. Most often, Aedes mosquitoes, particularly Aedes aegypti in tropical areas, spread the disease. Based on the typical symptoms, a wide differential diagnosis can be made for zika virus infection. Reverse transcriptase polymerase chain reaction (RT-PCR) screening enables a more accurate diagnosis. Although there is now no vaccination available, staying inside and avoiding mosquito bites will help you avoid a lot of diseases. Despite the rarity of occurrences of Zika virus sickness in the most populous nation in Africa, it is crucial to increase public knowledge in order to avoid any potential health issues.

Screening of P-35S, P-FMV, and T-NOS genetic elements in microwave-treated genetically modified cereal flours

Reliable and efficient methods for detecting genetically modified organisms (GMOs) in unprocessed and processed food will be essential for establishing an effective system for traceability all along the supply chain. It is important to understand the detection of GMOs following microwave treatment, which is a common processing method used in various food products such as flours. Therefore, this study aimed to detect the presence of Cauliflower mosaic virus (CaMV) 35S promoter (P-35S), Figwort mosaic virus (FMV) promoter (P-FMV), and T-NOS (nopaline synthase terminator) genetic elements in DNA samples from untreated and microwave-treated genetically modified (GM) cereal flour samples using the qualitative polymerase chain reaction (PCR) based screening method. DNA was extracted from all samples, and the efficiency of the qualitative PCR screening technique was tested by the verification studies. We performed an inhibition study with plant-specific actin (ACT) gene to the effectiveness of confirming the DNA extraction method. Then, we made the confirming of the qualitative PCR system by method performance testing criteria. The high quality and quantity of the DNA extracts from untreated and microwave-treated flour samples indicated the applicability of qualitative PCR screening assays. The results showed that microwave radiation does not significantly impact the genetic element screening in flour materials. Untreated and microwave-treated flour samples had amplifiable DNA for the simultaneous screening of three genetic elements. The qualitative screening tests conducted in this study produced dependable outcomes, thus, can be successfully used for monitoring in control laboratories.