Negative Polymerase Chain Reaction Research Articles

Heart Transplantation from COVID-19–Positive Donors: A Word of Caution

During the COVID-19 pandemic, efforts to maintain solid-organ transplantation have continued, including the use of SARS-CoV-2-positive heart donors. We present our institution's initial experience with SARS-CoV-2-positive heart donors. All donors met our institution's Transplant Center criteria, including a negative bronchoalveolar lavage polymerase chain reaction result. All but 1 patient received postexposure prophylaxis with anti-spike monoclonal antibody therapy, remdesivir, or both. A total of 6 patients received a heart transplant from a SARS-CoV-2-positive donor. One heart transplant was complicated by catastrophic secondary graft dysfunction requiring venoarterial extracorporeal membrane oxygenation and retransplant. The remaining 5 patients did well postoperatively and were discharged from the hospital. None of the patients had evidence of COVID-19 infection after surgery. Heart transplants from SARS-CoV-2 polymerase chain reaction-positive donors are feasible and safe with adequate screening and postexposure prophylaxis.

Prolonged Fever in a Child With Congenital Heart Disease and a Prosthetic Valve.

Prolonged Fever in a Child With Congenital Heart Disease and a Prosthetic Valve.

Monkeypox-Associated Central Nervous System Disease: A Case Series and Review.

Monkeypox virus (MPXV) disease has been declared a public health emergency by the World Health Organization, creating an urgent need for neurologists to be able to recognize, diagnosis, and treat MPXV-associated neurologic disease. Three cases of MPXV-associated central nervous system (CNS) disease occurring during the 2022 outbreak, and their associated imaging findings are presented, with 2 cases previously published in a limited capacity in a public health bulletin. Three previously healthy immunocompetent gay men in their 30s developed a febrile illness followed by progressive neurologic symptoms with presence of a vesiculopustular rash. MPXV nucleic acid was detected by polymerase chain reaction (PCR) from skin lesions of 2 patients, with the third patient having indeterminate testing but an epidemiologic link to a confirmed MPXV disease case. Cerebrospinal fluid demonstrated a lymphocytic pleocytosis, elevated protein, and negative MPXV-specific PCR. In 2 patients, magnetic resonance imaging of the brain and spine demonstrated partially enhancing, longitudinally extensive central spinal cord lesions with multifocal subcortical, basal ganglia, thalamic, cerebellar, and/or brainstem lesions. The third patient had thalamic and basal ganglia lesions. All patients received 14 days of tecovirimat, and 2 patients also received multiple forms of immunotherapy, including intravenous immunoglobulin, pulsed high-dose steroids, plasmapheresis, and/or rituximab. Good neurologic recovery was observed in all cases. MPXV can be associated with CNS disease. It is unclear whether this is from a parainfectious immune-mediated injury or direct CNS viral invasion. ANN NEUROL 2023;93:893-905.

Breath Analysis of COVID-19 Patients in a Tertiary UK Hospital by Optical Spectrometry: The E-Nose CoVal Study.

Throughout the SARS-CoV-2 pandemic, diagnostic technology played a crucial role in managing outbreaks on a national and global level. One diagnostic modality that has shown promise is breath analysis, due to its non-invasive nature and ability to give a rapid result. In this study, a portable FTIR (Fourier Transform Infra-Red) spectrometer was used to detect chemical components in the breath from Covid positive symptomatic and asymptomatic patients versus a control cohort of Covid negative patients. Eighty-five patients who had a nasopharyngeal polymerase chain reaction (PCR) test for the detection of SARS-CoV-2 within the last 5 days were recruited to the study (36 symptomatic PCR positive, 23 asymptomatic PCR positive and 26 asymptomatic PCR negative). Data analysis indicated significant difference between the groups, with SARS-CoV-2 present on PCR versus the negative PCR control group producing an area under the curve (AUC) of 0.87. Similar results were obtained comparing symptomatic versus control and asymptomatic versus control. The asymptomatic results were higher than the symptomatic (0.88 vs. 0.80 AUC). When analysing individual chemicals, we found ethanol, methanol and acetaldehyde were the most important, with higher concentrations in the COVID-19 group, with symptomatic patients being higher than asymptomatic patients. This study has shown that breath analysis can provide significant results that distinguish patients with or without COVID-19 disease/carriage.

Enhanced surveillance of notifications of hepatitis C to Queensland Health up to 19 years previously.

In this study we aimed to assess the utility of following up historical hepatitis C notifications for enhanced surveillance and linking cases to further testing and treatment. Queensland hepatitis C notifications from June 2018, 2013, 2008 and 2003 who were not incarcerated at the time of testing were followed up. The most recent identified clinicians for cases were contacted by telephone. When no information about a current clinician was available, the case was contacted via a letter or text message. Clinicians and cases were encouraged to pursue further testing and treatment and provide information about management. Following notification but prior to this study's follow-up, a majority of cases (309/532; 58%) had a negative polymerase chain reaction (PCR) test or underwent treatment.Clinician follow-up was successful in 21% of eligible cases, with the proportion decreasing with increasing time since notification. In conclusion, contacting clinicians to link notified cases to further testing and treatment may increase testing and treatment in a small proportion of cases notified up to nine years post-notification. From our experience, the follow-up of notifications before this time is unlikely to result in improved outcomes.

Upper Airway Mucociliary Clearance is Impaired in Dyspneic COVID-19 Patients.

Covid-19 is transmitted mainly by respiratory droplets and as the upper airway mucosa is the first innate immune barrier, it is crucial to understand the effects of SARS-CoV-2 on this system. In the current study, we aimed to evaluate the nasal mucociliary clearance in patients with SARS-CoV-2 infection and their symptom development. Observational cross-sectional study. The nasal mucociliary clearance (NMC) time was evaluated by the saccharin test and the results were compared between patients with SARS-CoV-2 infection (group 1) and controls (group 2, asymptomatic patients with a negative polymerase chain reaction test). We also compared the NMC time for each specific symptom suffered by participants in group 1 with the NMC time of the control group as well as with the patients in group 1 who were asymptomatic. There was a significant increase in NMC time in group 1with dyspnea when compared to the control group (p = 0.032) and also when compared to patients who were infected were not dyspneic (p = 0.04). There were no differences in the clearance times when considering other symptoms. COVID-19 patients with dyspnea present with altered nasal mucociliary clearance.

Diagnostic value of Treponema pallidum PCR test in real practice.

Syphilis is a sexually transmitted infection (STI) caused by the pathogen Treponema pallidum. Its incidence is increasing in our country, especially among men who have sex with men (MSM). Serological tests are still the most widely used technique for diagnosis. The need for an early diagnosis has prompted the introduction of fast techniques, such as Treponema pallidum detection by polymerase chain reaction (PCR) on mucocutaneous samples. The objective of this work is to analyse the sensitivity of this technique in a series of patients diagnosed with syphilis at our centre. Retrospective review of all cases diagnosed with syphilis at our centre between May 2017 and May 2021. A total of 203 cases of syphilis were diagnosed with serologic tests: 33% were primary syphilis and 53.1% secondary syphilis. PCR for Treponema pallidum was performed in 117 (57,6%) cases. The sensitivity was highest (95,2%) when performed on samples from mucocutaneous ulcers in primary syphilis. This value decreased to 69,4% in secondary syphilis, although there were variations between the types of samples. The PCR test has a high diagnostic value when performed on ulcer exudates in patients with primary syphilis. Its most relevant advantages in clinical practice are the possibility of an early diagnosis before serological tests during the window period, the ability to confirm reinfections in patients with persistent positivity of reaginic antibodies and a history of treated syphilis. Nevertheless, given that a negative PCR test may not rule out infection by Treponema pallidum, serologic tests are still necessary for everyday practice.

The Effect of Post-Coronavirus Disease 2019 Infection on the Retinal Microvasculature.

To evaluate the short-term microvasculature changes of the macula and optic disc following coronavirus disease 2019 (COVID-19). This study included 150 eyes (50 eyes of healthy controls and 100 eyes of patients) during the 1st month following COVID-19 recovery, as evidenced by two negative polymerase chain reactions. A complete ophthalmic examination and optical coherence tomography angiography were performed to detect the deep and superficial macular vessel density (VD). In addition, the VD of the optic disc was evaluated. Deep VD (DVD) showed a statistically significant decrease in post-COVID-19 patients, particularly those with severe COVID-19. This reduction occurred in the whole image, parafoveal, and perifoveal VD (P = 0.002, P = 0.002, and P < 0.001, respectively). Concerning the superficial VD (SVD), only the superior hemisphere of the whole image density was statistically significantly reduced (P = 0.037). There was no statistically significant difference in foveal VD (both deep and superficial vessel) among the study groups (P = 0.148 and P = 0.322, respectively). Regarding the foveal avascular zone (FAZ), there was no statistically significant among groups (P = 0.548). Regarding the optic disc, the whole image VD and redial peripapillary capillary VD demonstrated a highly significant decrease, particularly in cases of severe COVID-19. Conversely, inside disc VD showed a nonsignificant change among the study groups. According to the findings of the current study, retinal microvasculature was affected in the 1st month following recovery from COVID-19. DVD was significantly reduced more than SVD. In addition, peripapillary VD decreased, whereas the FAZ was unaffected.

A serum interleukin-6-based analysis of patients with nasopharyngeal swab reverse-transcriptase polymerase chain reaction negative for SARS-COV-2 infection with CO-RADS 4 and 5 on computed tomography of the chest

Background: Serum interleukin 6 (IL-6) levels have been studied in the diagnostic evaluation of patients with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) disease (COVID-19).Methods: We studied the utility of treatment with tocilizumab in COVID-19 patients (n=19) with a negative nasopharyngeal swab real time reverse transcriptase polymerase chain reaction (RT-PCR) test for SARS-CoV-2 who had suggestive computed tomography (CT) findings, namely, COVID-19 Reporting and Data System (CO-RADS) 4,5.Results: Receiver operator characteristic (ROC) curve analysis showed that serum IL 6 at a cut-off of >56.9 pg/L was a predictor of mortality in nasopharyngeal swab RT-PCR negative patients with suggestive CT findings. Tocilizumab had no significant effect on the mortality.Conclusions: In nasopharyngeal swab RT-PCR negative patients with suggestive chest CT findings, elevated serum IL-6 levels > 56.9 pg/L predicted mortality. However, treatment with tocilizumab had no effect on mortality.

Thrombosis Occurrence in COVID-19 Compared With Other Infectious Causes of ARDS: A Contemporary Cohort.

Thrombosis occurrence in coronavirus disease 2019 (COVID-19) has been mostly compared to historical cohorts of patients with other respiratory infections. We retrospectively evaluated the thrombotic events that occurred in a contemporary cohort of patients hospitalized between March and July 2020 for acute respiratory distress syndrome (ARDS) according to the Berlin Definition and compared those with positive and negative real-time polymerase chain reaction results for wild-type severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) using descriptive analysis. The association between COVID-19 and thrombotic risk was evaluated using logistic regression. 264 COVID-19-positive (56.8% male, 59.0 years [IQR 48.6-69.7], Padua score on admission 3.0 [2.0-3.0]) and 88 COVID-19-negative patients (58.0% male, 63.7 years [51.2-73.5], Padua score 3.0 [2.0-5.0]) were included. 10.2% of non-COVID-19 and 8.7% of COVID-19 patients presented ≥ 1 clinically relevant thrombotic event confirmed by imaging exam. After adjustment for sex, Padua score, intensive care unit stay, thromboprophylaxis, and hospitalization length, the odds ratio for thrombosis in COVID-19 was 0.69 (95% CI, 0.30-1.64). We, therefore, conclude that infection-induced ARDS carries an inherent thrombotic risk, which was comparable between patients with COVID-19 and other respiratory infections in our contemporary cohort.

Long-term self-reported health and disability after COVID-19 in public employees

BackgroundAlthough the symptomatology has been assessed in multiple studies among persons recovering from coronavirus disease 2019 (COVID-19), less is known regarding long-term general health and disability. We aimed to assess long-term self-reported disability in public employees after predominantly mild COVID-19 in comparison with individuals who had negative COVID-19 polymerase chain reaction (PCR) test results.MethodsPublic employees within Region Västra Götaland were offered tests to identify SARS-CoV-2 infection (n = 56,221) and were invited to complete an online survey that included the World Health Organization Disability Assessment Schedule. Questionnaires were sent out between January 26 and March 5, 2021. A total of 14,222 (25.3%) employees responded, of which 10,194 (18%) were included (women n = 8749, 85.8%). Of these, 7185 (70.5%) participants had a negative PCR test result (controls). A total of 1425 (14%) had a positive PCR result and were categorized in the sub-acute phase (4–12 weeks post COVID-19), and 1584 (15.5%) had a positive PCR test and were categorized in the post COVID-19 phase (> 12 weeks).ResultsFifty-two percent of controls rated disability of varying degrees, versus 73% and 64% of participants in the sub-acute and post COVID-19 phase, respectively. Being “emotionally affected” was the most frequently reported disability in the sub-acute phase, the post COVID-19 phase, as well as in controls. The proportion of participants reporting difficulties for 20–30 days was higher in the sub-acute phase than in the post COVID-19 phase (27.9% vs. 21.8%, p < 0.001) as well as in a comparison between participants in the post COVID-19 phase and controls (21.8% vs 14.2%, p < 0.001). Compared with controls, severe disability was more common in the post COVID-19 phase among both women (15.8% vs. 10.7%,), and men (9.8% vs. 6.8%).ConclusionsDisability was present in all groups; however, reported disability was greater in the sub-acute phase than in the post COVID-19 phase. The higher levels of disability reported in the COVID-19 patient population may indicate a persisting need for rehabilitation and recovery. In general, women reported a greater degree of disability than men in the sub-acute and post COVID-19 phases.

The relationship between cardiological parameters and PCR in patients with coronavirus infection: A cross-sectional study

Cardiovascular injuries induced by SARS CoV-2 have been reported repeatedly in various studies. Therefore, it is necessary to understand cardiac complications at a low cost, quickly. This study aimed to determine the relationship between cardiological parameters and polymerase chain reaction (PCR) in patients with coronavirus infection. : Patients who were admitted to the emergency department due to the ongoing pandemic, all patients with similar symptoms to coronavirus disease 2019 infection were initially admitted to the respiratory emergency room and underwent subsequent evaluations to confirm or rule out SARS-COV2 infection symptoms were assessed for eligibility. Patient were categorized into 2 groups 1. Positive PCR and negative PCR groups. Binary logistic regression was performed to assess the effect of several factors on the likelihood of developing positive troponin, reduced ejection fraction (EF), and Positive brain natriuretic peptide (BNP). Among 195 patients included, 115 (58.9%) had positive PCR. Patient in the positive PCR and negative PCR were 58.04 ± 18.03 and 59.19 ± 15.38 years of age, respectively. Patients in the "positive PCR" were significantly less likely to have chronic kidney disease (6.69% vs 17.5%, P value: .022), consume calcium channel blockers (6.69% vs 18.75%, P value:0.012). At the univariable level, positive PCR was significantly associated with fewer odds for positive BNP (OR:0.46, P = .019); nevertheless, the association was no longer significant after adjusting for confounders (adjusted OR:0.56, P = .158). Unadjusted positive PCR results were not found to have a significant association with positive troponin or reduced EF. Likewise, multivariable regression revealed no association between positive PCR and positive troponin (aOR:1.28, P = .529) and reduced EF (aOR:0.65, P = .369). PCR positivity did not result in positive troponin and BNP and did not appear to decrease EF. In other words, serial troponin and BNP checks and initial echocardiography in coronavirus disease 2019 respiratory emergencies do not make significant differences in diagnostic and therapeutic management and inpatient outcomes of patients with positive or negative PCR and are not specific findings. Evidence suggests some coronavirus-induced cardiac complications will be manifested in the long term.

Estimation of the incubation period and generation time of SARS-CoV-2 Alpha and Delta variants from contact tracing data.

Quantitative information on epidemiological quantities such as the incubation period and generation time of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants is scarce. We analysed a dataset collected during contact tracing activities in the province of Reggio Emilia, Italy, throughout 2021. We determined the distributions of the incubation period for the Alpha and Delta variants using information on negative polymerase chain reaction tests and the date of last exposure from 282 symptomatic cases. We estimated the distributions of the intrinsic generation time using a Bayesian inference approach applied to 9724 SARS-CoV-2 cases clustered in 3545 households where at least one secondary case was recorded. We estimated a mean incubation period of 4.9 days (95% credible intervals, CrI, 4.4-5.4) for Alpha and 4.5 days (95% CrI 4.0-5.0) for Delta. The intrinsic generation time was estimated to have a mean of 7.12 days (95% CrI 6.27-8.44) for Alpha and of 6.52 days (95% CrI 5.54-8.43) for Delta. The household serial interval was 2.43 days (95% CrI 2.29-2.58) for Alpha and 2.74 days (95% CrI 2.62-2.88) for Delta, and the estimated proportion of pre-symptomatic transmission was 48-51% for both variants. These results indicate limited differences in the incubation period and intrinsic generation time of SARS-CoV-2 variants Alpha and Delta compared to ancestral lineages.

Report of the first case of Pediatric Multisystemic Inflammatory Syndrome at Hospital de Especialidades Jóse Carrasco Arteaga in 2020

BACKGROUND: PIMS/MIS-C involves an innate immune response of the host against a trigger, in this case a virus, which causes hyperinflammation induced by a cytokine storm; this immune response is responsible for the systemic clinical manifestations and cardiac complications. The syndrome has been diagnosed usually between 2 and 6 weeks after infection by SARS-CoV-2; however, many patients have negative PCR (polymerase chain reaction) tests for the virus and positive immunoglobulin tests, suggesting that the syndrome appears in the early stages of convalescence from COVID-19. CASE REPORT: A 15-year-old female patient, previously healthy, with a history of close contact with a relative with COVID-19 confirmed by PCR 15 days before the onset of symptoms. He presented a 5-day history of fever, vomiting, and diarrheal stools; 48 hours later she presented with generalized asthenia, hyporexia and epistaxis. We evidenced elevation of immunoglobin G against SARS-COV-2 and negative PCR. In complementary tests, we evidenced elevation of inflammatory markers (CRP, procalcitonin, ferritin, D-dimer) and BNP; the echocardiogram revealed a condensation process in the pericardium, as well as coronary ectasia. Evolution: After confirming the diagnosis of PIMS/MIS-C, we started treatment with immunoglobulin, prednisone, and acetylsalicylic acid. On the fourth day of hospitalization, she presented diaphoresis and moderate pain in the epigastric region that did not subside; Abdominal CT revealed signs of mesenteric adenitis, which reconfirmed the proposed diagnosis. After completing 72 hours of treatment, and being evaluated again, she was discharged. We prescribed continuance of prednisone treatment and acetylsalicylic acid and 500 milligrams of amoxicillin orally every 8 hours. In addition, we recommended follow-up by the cardiology service one week after hospital discharge. The patient remains without complications after this episode. CONCLUSION: The diagnostic suspicion of PIMS/MIS-C must be present in all patients with characteristic symptoms, under the age of 20 years, with a history of contact with a positive COVID-19 patient and who in most cases will have a positive PCR test or positive serology. Given the multiple and variable clinical manifestations, it is important to take into account the differential diagnoses, especially Kawasaki Disease, whose clinical manifestations this syndrome mimics. Management and treatment depend on the patient's condition, but treatment with intravenous immunoglobulin is generally recommended; most patients respond to a single dose, as in the present case.

1287: METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS NARES SCREEN LEADS TO VANCOMYCIN DE-ESCALATION IN TRAUMA

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) polymerase chain reaction (PCR) nares swab is a proven screening tool to de-escalate vancomycin therapy due to its high negative predictive value in pneumonia and other infections. However, data are lacking on the utility of this test in the critically ill trauma population, which is at high risk for hospital-acquired infections. This study evaluated the impact of empiric MRSA PCR nares screening on de-escalation of vancomycin therapy in critically ill trauma patients. Methods: This is a single center, pre-post observational cohort study conducted at an academic, level I trauma center. Adult patients who were admitted for trauma and received empiric vancomycin therapy for suspected infection in the trauma intensive care unit were included. Patients were grouped by whether they were included 6 months before (pre-cohort) and 6 months after (post-cohort) the implementation of empiric rapid MRSA PCR nares screening in September 2021. The primary outcome was duration of vancomycin therapy. Secondary outcomes included rate of acute kidney injury (AKI), positive MRSA screen or cultures, ICU length of stay (LOS), hospital LOS, and mortality. Results: One-hundred fifty-eight patients were included: 82 in the pre-cohort group and 76 in the post-cohort group. Ninety vancomycin initiation orders were observed in the pre-cohort versus 80 orders in the post-cohort. Baseline demographics were similar between groups. There was a significantly higher rate of MRSA PCR orders in the post-cohort group (75%) compared to (21%) in the pre-cohort (p< 0.001). The pre-cohort group had higher rates of positive MRSA screens (26 vs. 13%, p=0.18) and MRSA positive cultures (17 vs. 9%, p=0.22) compared to post-cohort. There were no reported instances of positive MRSA cultures with a negative PCR. Vancomycin duration was significantly shorter in the post-cohort group (2.55 days vs. 3.5, p=0.02). Hospital and ICU LOS, rates of AKI, and mortality were not different between groups. Conclusions: Implementation of empiric rapid MRSA nares screening led to a shorter overall duration of vancomycin therapy by nearly one day in critically ill trauma patients. Further study is required to evaluate the potential cost savings of empiric MRSA screening in critically ill trauma patients.

1804. Quality Improvement Project to improve empiric antibiotic therapy for skin and soft tissue infections using the nasal MRSA PCR

Abstract Background Large cohort studies from the Veteran’s Administration support the negative predictive value of the methicillin resistant Staphylococcus aureus (MRSA) nasal polymerase chain reaction (PCR) swabs for skin and soft tissue infection (SSTI). We investigated the use of the nasal MRSA PCR swab to reduce the inappropriate empiric use of vancomycin and piperacillin-tazobactam for SSTI at our institution. Methods Between July and August 2021, we educated Hospital Internal Medicine (HIM) and Internal Medicine Residents (IMR) on the basics of SSTI management and the utility of a negative nasal MRSA PCR swab. Data on empiric use of vancomycin and piperacillin-tazobactam for SSTIs were collected 3/1/2021 to 7/1/2021 (pre-intervention) and 8/30/2021 to 12/29/2021 (post-intervention). We excluded patients who didn’t have an SSTI, weren’t managed by IMR or HIM, and whose empiric antibiotics were for another indication. To account for antibiotics started in the emergency department (ED), a patient was regarded as having received appropriate empiric antibiotics if they were de-escalated in the first 24 hours of being admitted to IMR or HIM. Example of education material. These fliers were distributed to resident rooms following a series of noon conferences Results Pre-intervention there were 106 patient encounters. In 56 of these (52.8%), patients were placed on inappropriate empiric antibiotics. Post-intervention, there were 72 patient encounters. In 23of these (31.9%), patients were placed on inappropriate empiric antibiotic for an absolute risk reduction (ARR) of 20.9% with a statistically significant 95% CI (6.522 - 35.249) and a number needed to treat (NNT) of 4.78. Nasal MRSA PCR testing increased from 40.6% to 56.9%. The ARR among the HIM teams was 21.3% and was statistically significant with a 95% CI (4.24 - 38.394). Their proportion of MRSA testing increased from 41.5% to 50%. The IMR teams had an ARR of 14.8%, which was not statistically significant (95% CI, -11.339 to 40.884). Their MRSA nasal PCR testing increased from 37.5% to 72%. Conclusion Education on SSTI treatment coupled with use of the MRSA nares PCR appeared to improve empiric antibiotic choices. Residents stated that the nasal MRSA PCR facilitated empiric antibiotic choice. Positive ARR was seen in whole and sub-group analyses of physician teams. We theorize that the lack of statistical significance in the IMR group was related to a small sample size. Disclosures All Authors: No reported disclosures.

85. Plasma Cell Free Deoxyribonucleic Acid (cfDNA) Fungal Polymerase Chain Reaction (PCR): a Non-Invasive Diagnostic for Invasive Fungal Disease Evaluation in At-Risk Pediatric Oncology and Stem Cell Transplant Patients

Abstract Background Molecular diagnostics appear promising for early, non-invasive detection of invasive fungal disease (IFD) in immunocompromised patients. Our clinical lab developed and validated cell free DNA (cfDNA) fungal polymerase chain reaction (PCR) assays (Table 1), which have been in clinical use since November 2020 and were recently included in an institutional pediatric clinical care pathway for prolonged febrile neutropenia. We aimed to evaluate the performance of these plasma cfDNA fungal PCR assays in pediatric oncology and hematopoietic stem cell transplant (HSCT) patients with clinical concern for IFD. Methods We initiated an observational study of inpatient oncology and HSCT patients who had plasma mold panel (+/- Candida panel) cfDNA fungal PCR obtained as part of an IFD evaluation at our freestanding quaternary care children’s hospital. The primary outcome was IFD clinical diagnosis (proven, probable, possible, unlikely) as per EORTC/MSG† definitions within one month of the cfDNA fungal PCR, which was assigned independently by two physicians, with a third physician utilized in cases of discrepancy. Patient demographics, hospital course, imaging results, and clinical laboratory data were abstracted and maintained in an encrypted REDCap© database. Results In a preliminary analysis of October 2021-March 2022 data, there were 21 IFD evaluations for 18 patients (Table 2). Most oncology evaluations were for prolonged febrile neutropenia, while many HSCT were non-neutropenic, but on enhanced immunosuppression with new clinical concerns (e.g., respiratory symptoms, persistent fever). Plasma cfDNA detected a mold or yeast consistent with the clinical presentation in 100% of the five proven/probable cases (Figures 1 &amp; 2). All 14 possible IFD cases had a negative cfDNA fungal PCR. Proven cases are designated with blue icon, while the probable case is orange. All five cases were in oncology patients who did not have history of hematopoietic stem cell transplant. Cell free DNA (cfDNA) fungal polymerase chain reaction (PCR) results include the organism identified, followed by the cycle threshold (Ct) in parenthesis. Maximum Ct for the assay is 45. Abbreviations: AG: Aspergillus galactomannan index (Ref &amp;lt;0.5); ALL: acute lymphoblastic leukemia. BAL: bronchoalveolar lavage; 1,3-BDG: 1,3-beta-D-glucan (Ref &amp;lt;60 pg/ml); cfDNA: cell free deoxyribonucleic acid; CNS: central nervous system; EORTC/MSG: European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group &amp; the National Institute of Allergy &amp; Infectious Diseases Mycoses Study Group; L-AmB: liposomal amphotericin B; SARS-CoV-2: severe acute respiratory syndrome associated with coronavirus 2. Conclusion Upfront plasma cfDNA fungal PCR successfully detected a relevant mold or yeast with short turnaround time in 100% of cases that were later classified as proven/probable. This appears promising for early, noninvasive diagnosis that enables prompt optimization of antifungal and surgical management, particularly for mucormycosis cases. Additional data may permit consideration of Mucorales agents PCR as a new EORTC/MSG mycologic criteria. Disclosures Lauren E. Kushner, MD, Roman: Client of my husband's business..

Rapid diagnosis of intra-amniotic infection using nanopore-based sequencing.

Early diagnosis and treatment of intra-amniotic infection is crucial. Rapid pathogen identification allows for a definite diagnosis and enables proper management. We determined whether the 16S amplicon sequencing performed by a nanopore sequencing technique make possible rapid bacterial identification at the species level in intra-amniotic infection. Five cases of confirmed intra-amniotic infection, determined by either cultivation or 16S rDNA polymerase chain reaction (PCR) Sanger sequencing, and 10 cases of women who underwent mid-trimester genetic amniocentesis were included. DNA was extracted from amniotic fluid and PCR was performed on the full-length 16S rDNA. Nanopore sequencing was performed. The results derived from nanopore sequencing were compared with those derived from cultivation and Sanger sequencing methods. Bacteria were successfully detected from amniotic fluid using nanopore sequencing in all cases of intra-amniotic infection. Nanopore sequencing identified additional bacterial species and polymicrobial infections. All patients who underwent a mid-trimester amniocentesis had negative cultures, negative 16S PCR Sanger sequencing and nanopore sequencing. Identification of the microorganisms using nanopore sequencing technique at the bacterial species level was achieved within 5-9h from DNA extraction. This is the first study demonstrating that the nanopore sequencing technique is capable of rapid diagnosis of intra-amniotic infection using fresh amniotic fluid samples.

Presence of SARS-CoV-2 on the conjunctival mucosa in patients hospitalized due to COVID-19: Pathophysiological considerations and therapeutic implications.

Coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) resulted in a worldwide pandemic, due to its great capacity to invade the human body. Previous studies have shown that the primary route of invasion of this virus is the human respiratory tract via the co-expression of ACE2 receptor and TMPRSS2, a serine protease on the cellular surface. Interestingly, this condition is present not only on the respiratory epithelium but on the conjunctival mucosa, as well. Thus, we hypothesized that SARS-CoV-2 is present on the conjunctival mucosa. To prove that SARS-CoV-2 can be detected in the conjunctiva. Previously nasopharyngeal swab-sample based real-time polymerase chain reaction (PCR) positive COVID-19 infected patients were selected at the COVID Care Centers of Semmelweis University, Budapest, Hungary. The study was approved by the ethical committee of Semmelweis University. During their recovery, both nasopharyngeal and conjunctival swab-samples were taken and PCR method was used to detect the presence of SARS-CoV-2 RNA. Appropriate statistical analysis was performed. The study population consisted of 97 patients, 49 females (50.5%) and 48 males (49.5%), with a mean age of 67.2 ± 11.9 years. During recovery, with nasopharyngeal swabs, the PCR test was positive in 55 cases (56.70%), whereas with conjunctival swabs it was positive in 8 cases (8.25%). Both tests were positive in 5 cases (5.15%). In some patients, ocular symptoms were observed as well. The rest of the patients (29 cases) had negative nasopharyngeal PCR tests during recovery. Although only in few cases, the data of the present study provides a proof of concept that SARS-CoV-2 can be present on the conjunctival mucosa even in nasopharyngeal negative patients, a finding, which can have clinical importance. Also, on the basis of these findings one can hypothesize that - in addition to the respiratory tract - the conjunctiva can be an entrance route for SARS-CoV-2 to the human body. Thus, in high-risk conditions, in addition to covering the mouth and nose with mask, the protection of the eyes is also strongly recommended.

Detectable Antibodies in Newborn Cord Blood Following Maternal COVID-19 Infection Before Pregnancy: A Case Report

: In this report, we present a case of an infant with detectable antibodies in the umbilical cord blood who was born to a healthcare professional mother diagnosed with coronavirus disease 2019 (COVID-19) before pregnancy and recovered, and also discuss the antibody level in light of relevant literature. Data consisted of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) polymerase chain reaction (PCR) test and negative pre-pregnancy repeat real-time polymerase chain reaction (RT-PCR) test of a 29-year-old medical doctor mother, follow-up antibody test results in pregnancy, and antibodies detected in the cord blood of the newborn after birth. The study data were collected through face-to-face interviews with the mother. In conclusion, this case shows that SARS-CoV-2 immunoglobulin G (IgG) antibodies can be detected in the baby’s cord blood immediately after birth from a mother who was diagnosed with COVID-19 before becoming pregnant and subsequently recovered with pharmacological treatment.