Polymerase Chain Reaction Test Research Articles

Laboratory detection of donors implicated in transfusion-transmitted malaria.

Transfusion-transmitted malaria (TTM) is rare in non-endemic areas (non-EAs) but can potentially be fatal. This review analyzes the laboratory results of donors causing TTM in non-EAs, to assess the detectability of their Plasmodium infection by molecular or antibody tests. TTM cases in the United States, Canada, and Europe since 2010 were identified through a literature review. Authors and laboratories were contacted for missing details about sample types and laboratory methods. Results of Plasmodium polymerase chain reaction (PCR) and antibody tests were summarized. Twelve cases of TTM and one bone marrow transplant transmission were identified. Of the 13 source donors, 12 were tested by PCR, 10 were positive on at least one sample; the 2 negative donors were tested only on retained segments of blood refrigerated for several weeks. All donors were PCR positive on a fresh sample except one who was positive on a retained but not a fresh sample. These PCRs targeted Plasmodium DNA with sensitivities in the range of 1000-10,000 parasites/mL. Antibody EIA was positive in only three of seven donors tested. This review found that antibody EIAs failed to detect four of the seven TTM donors tested. DNA-based PCRs were able to detect Plasmodium infection in all donors tested except for two tested only on samples likely to have deteriorated from prolonged storage. Recently developed ribosomal RNA-based molecular donor screening assays are approximately 1000 fold more sensitive than these DNA-based PCRs, holding promise as a potential method to further reduce TTM.

Utilization of Syndromic Vaginitis Diagnostic Testing Reduces 6-Month Follow-Up Outpatient Service Healthcare Costs—A Real-World Data Analysis

Background/Objectives: Vaginitis is a common infection among women of reproductive age. Although various diagnostic methodologies exist, diagnosis without the utilization of available diagnostic tests remains prevalent. This study aimed to assess downstream healthcare utilization and the cost of patients with and without diagnostic testing. Methods: This retrospective, observational study utilized the IQVIA PharMetrics® Plus database from July 2020 to October 2023. Patients with an index claim (ICD-10 code indicating vaginitis) were categorized into two cohorts: those who received a syndromic polymerase chain reaction (PCR) test and those who had no documented test on the index date or within two days. Total and service-specific healthcare resource utilization and costs were assessed for 6 months following the index event. This study was designed to inform how Syndromic Vaginitis PCR testing is used to make treatment decisions and to track outpatient and inpatient healthcare utilization for 6 months post index date represented by cost. Results: Patients who received a Syndromic Vaginitis PCR test had significantly fewer outpatient medical services in the 6 months following initial diagnosis compared to those who received no diagnostic test. This was largely attributed to a substantial decrease in other medical service visits, resulting in mean cost savings of USD 2067 (Syndromic PCR = USD 6675, SD = USD 17,187; No Test = USD 8742, SD = USD 29,894) (p-value 0.0009). Conclusions: Many vaginitis patients do not receive testing, but Syndromic Vaginitis PCR testing may be an effective diagnostic tool for reducing costs associated with vaginitis infections.

Noninvasive Early Detection of Systemic Inflammatory Response Syndrome of COVID-19 Inpatients Using a Piezoelectric Respiratory Rates Sensor

In 2020, 20% of patients with COVID-19 developed severe complications, including life-threatening pneumonia with systemic inflammatory response syndrome (SIRS). We developed a preliminary SIRS monitor that does not require blood sampling, is noninvasive, and can collect data 24 h per day. The proposed monitor comprises a piezoelectric respiratory sensor located beneath the patient’s mattress and a fingertip pulse sensor that determines ultra-high accuracy respiratory rate (mode of a 40-min frequency distribution of respiratory rates (M40FD-RR)). We assessed the clinical performance of the M40FD-RR preliminary SIRS monitor in 29 patients (12 female, 17 male, aged 15–90 years) hospitalized at Suwa Central Hospital with COVID-19, which was confirmed by a positive polymerase chain reaction test. SIRS was evaluated by logistic regression analysis using M40FD-RR, heart rate, age, and sex as explanatory variables. We compared the results of 109 examinations of 29 COVID-19 inpatients with SIRS against those determined by the proposed monitor. The proposed monitor achieved 75% sensitivity and 83% negative predictive value, making it a promising candidate for future 24 h noninvasive preliminary SIRS tests.

Fatal borealpox in an immunosuppressed patient treated with antivirals and vaccinia immunoglobulin - Alaska, 2023.

Borealpox virus (BRPV, formerly known as Alaskapox virus) is a zoonotic member of the Orthopoxvirus genus first identified in a person in 2015. In the six patients with infection previously observed BRPV involved mild, self-limiting illness. We report the first fatal BRPV infection in an immunosuppressed patient. A man aged 69 years from Alaska's Kenai Peninsula was receiving anti-CD20 therapy for chronic lymphocytic leukemia. He presented to care for a tender, red papule in his right axilla with increasing induration and pain. The patient failed to respond to multiple prescribed antibiotic regimens and was hospitalized 65 days postsymptom onset for progression of presumed infectious cellulitis. BRPV was eventually detected through orthopoxvirus real-time polymerase chain reaction testing of mucosal swabs. He received combination antiviral therapy, including 21 days of intravenous tecovirimat, intravenous vaccinia immunoglobulin, and oral brincidofovir. Serial serology was conducted on specimens obtained posttreatment initiation. The patient's condition initially improved with plaque recession, reduced erythema, and epithelization around the axillary lesion beginning one-week post-therapy. He later exhibited delayed wound healing, malnutrition, acute renal failure, and respiratory failure. He died 138 days postsymptom onset. Serologic testing revealed no evidence the patient generated a humoral immune response. No secondary cases were detected. This report demonstrates that BRPV can cause overwhelming disseminated infection in certain immunocompromised patients. Based on the patient's initial response, early BRPV identification and antiviral therapies might have been beneficial. These therapies, in combination with optimized immune function, should be considered for patients at risk for manifestations of BRPV.

Role of admission rapid antigen testing (RATs) for COVID-19 on patients transferred from acute hospitals to a post-acute rehabilitation setting.

Rapid antigen tests (RATs) are suitable for point-of -care testing, require no laboratory time and give immediate results. However, are RATs useful for detecting asymptomatic COVID-19 infection when compared with polymerase chain reaction (PCR) testing in healthcare settings? The aim of this study was to implement a reliable testing system utilising RATs to promptly detect COVID-19 infection in predominantly asymptomatic patients transferred from acute hospitals to a post-acute rehabilitation unit (PARU). RAT testing was carried out on all new admissions without a history of confirmed Covid-19 infection within three months of admission. PCR testing was carried out on all patients with a positive RAT for confirmation purposes. The cycle threshold (Ct) values of COVID-19 detected results on PCR testing were examined to determine the utility of the RATs. A total of 1,403 patients were transferred to the PARU from January to December 2023. The results of the study revealed an 85% accuracy of RATs with a 15% rate of false negative results at the time of admission. All patients that had a positive RAT at the time of admission also had a positive PCR test. This testing algorithm resulted in early detection and prompt isolation of positive cases reducing the likely spread of COVID-19 infection, hospital outbreaks and bed/ward closures.

Prognostic value of serum amyloid A protein as a biomarker in the diagnosis of 2019 novel coronavirus disease (COVID-19)

BackgroundThe current study aimed to evaluate the prognostic value of serum amyloid A protein )SAA( protein as a biomarker in diagnosing 2019 novel coronavirus disease )COVID-19( infection. MethodsThe study was conducted on 123 patients with definitive COVID-19 infection referred to Shahid Beheshti and Sina hospitals in Hamedan province, Iran. Five-milliliter blood samples were taken from all included patients and serum was isolated using a centrifuge at 10,000 rpm for 10 minutes. Laboratory tests were conducted, including c-reactive protein (CRP), Erythrocyte Sedimentation Rate (ESR), potassium level, sodium blood test, platelets (PLT), complete blood count (CBC), lymphocyte count, and neutrophil count. The SAA enzyme-linked immunosorbent assay (ELISA) Kit was applied to measure the SAA level in serum samples. Results123 patients included 73 males and 50 females, age ±50. Sixty-six (53.7%) patients had negative CRP while 80 (65%) patients had normal ESR. Potassium levels were not normal among 111 (94.9%) patients. Seventy-seven (63.1%) patients had normal CBC, while 108 (87.8%) patients had neutrophils above the normal range. 94 (97.9%) patients over the age of 50 were positive for SAA. In terms of gender, men were the most frequent patients with SAA. There was a statistically significant relationship between the serum level of SAA and outcomes of patients with COVID-19 (p = 0.0001). 94% of patients with SAA ≤50 were recovered from COVID-19 infection. The sensitivity rate of SAA compared to polymerase chain reaction (PCR) and computed tomography scan (CT scan) tests was 93% and 99%, respectively. Moreover, the accuracy of SAA compared to PCR and CT scan tests was 52% and 96%, respectively. ConclusionResults indicate the SAA is a sensitive, but not specific biomarker in the early detection of COVID-19. The quantitative levels of SAA can be useful in predicting treatment outcomes among patients with COVID-19.

Relation Between Monocyte-to-lymphocyte Ratio and Depressive Symptoms in Patients with Non-severe COVID-19 Infection

Background and Aim: Mental health of non-hospitalized patients and those with non-severe infections has attracted lower attention in comparison to other patients. Circulating monocytes are deeply involved in all stages of COVID-19 infection. The present study aimed to investigate the relationship between monocyte-to-lymphocyte ratio (MLR) and depressive symptoms in patients with non-severe COVID-19 infection. Methods: The study included 312 patients with non-severe COVID-19 infection diagnosed on the basis of a positive reverse-transcriptase polymerase chain reaction (RT-PCR) test of nasopharyngeal swabs. Depressive symptoms were assessed using the validated Arabic version of the 7-item Hamilton Depression Rating Scale (HAMD). According to the obtained scores, patients were classified to have mild (10-13), moderate (14-17), or severe depression (>17). Results: The present study included 312 patients with non-severe COVID-19. According to HAMDS, clinically significant depression was diagnosed in 144 patients (46.2 %). They comprised 38 patients (12.2 %) with mild depression, 30 patients (9.6 %) with mild-tomoderate depression and 76 patients (24.4 %) with moderate-to-severe depression. Multivariate logistic regression analysis identified male sex [OR (95% CI): 2.07 (1.27-3.36), p = 0.003], presence of dyspnea [(OR (95 % CI): 1.99 (1.21-3.27), p = 0.007], D dimer levels [OR (95% CI): 2.32 (1.19-4.52), p = 0.013], MLR [OR (95% CI): 0.52 (0.28-0.99), p = 0.046] and abnormal CT findings [OR (95% CI): 1.79 (1.08-2.95), p = 0.023] as significant predictors of depression in the studied patients. Conclusion: Low MLR is related to depressive symptoms in patients with non-severe covid-19 infection. Other predictors include male sex, dyspnea, abnormal CT findings and elevated D-dimer levels.

Third cranial nerve palsy with confirmed HHV-6 positivity

We present a rare case of a 38-year-old man diagnosed with third cranial nerve paralysis, where no abnormalities were detected on MRI. Human Herpes Virus-6 (HHV-6) infection affecting the central nervous system and associated with cranial nerve paralysis is an unusual occurrence. This report underscores the importance of considering HHV-6 as a differential diagnosis in patients presenting with cranial nerve palsy and suggests the use of cerebrospinal fluid (CSF) HHV-6 polymerase chain reaction (PCR) testing for confirmation.

Toxoplasmosis infection in an HIV-negative patient presenting with clinical and MRI findings similar to those of multiple sclerosis

Toxoplasmosis is a parasitic infection that can present in various clinical forms, ranging from asymptomatic to severe neurological manifestations. The primary sources of infection include undercooked meat, unwashed produce and contact with cat faeces. Toxoplasmosis can lead to encephalitis, particularly in immunocompromised patients, and is often misdiagnosed as other neurological conditions such as multiple sclerosis (MS). We report the case of a 44-year-old male from Almaty, Kazakhstan, who presented with neurological symptoms including headaches, dizziness, diplopia, leg weakness and elevated blood pressure. The patient had a history of consuming undercooked meat, but no prior neurological conditions. Initial magnetic resonance imaging (MRI) revealed demyelinating lesions, leading to a diagnosis of MS. However, high levels of IgG antibodies against Toxoplasma gondii were detected, prompting further testing. A polymerase chain reaction test for toxoplasmosis was negative, but the patient was treated empirically with trimethoprim and sulfamethoxazole for six months. A follow-up MRI showed a significant reduction in brain lesions, and the patient’s symptoms improved.

Bartonella endocarditis: lessons learnt from an African cohort

Abstract Background Bartonella (B) quintana endocarditis is a preventable, vector-borne, and important cause of blood culture-negative endocarditis (BCNE). The ESC guideline for the management of endocarditis recommends blood serology (IgG titer ≥800) and polymerase chain reaction (PCR) testing on both blood and valvular tissue for diagnosis. Access to these molecular technologies is limited in Africa and there is a paucity of B. quintana endocarditis research on the continent. Purpose Record the clinical picture and explore the risk factors for disease and the diagnostic utility of immunofluorescence (IFA) in PCR-proven cases with B. quintana endocarditis. Methods Data were analysed from a prospective observational cohort study of patients with infective endocarditis managed at our institution. All BCNE cases from October 2019 to January 2024 were evaluated to identify patients with PCR-proven B. quintana on valvular tissue. A commercially available IFA was used for blood serology testing. 16S rRNA PCR testing with sequencing was performed on valvular tissue. Results Thirty five of the 58 cases (60.3%) with BCNE were diagnosed with Bartonella endocarditis. A diagnosis of PCR-proven B. quintana endocarditis was made in 21 cases. Of these, seven patients (33.3%) were either homeless or lived in informal housing. Thirteen patients (61.9%) had an alcohol-use disorder. Four patients (19%) were Human Immunodeficiency (HIV) virus- positive. Eighteen patients (85.7%) reported the onset of symptoms more than two weeks before presentation. Common presenting symptoms were dyspnea (n=21;100%), loss of weight (n=10;47.6%) and fever (n=7;33.3%). Common clinical features were digital clubbing (n=17;80.9%), pallor (n=13; 61.9%), microscopic haematuria (n=12; 57.1%) and severe regurgitant lesions (n=21; 100%). Blood serology was available in 17 patients with an IgG titer (1≥256) in all patients apart from one (1:64). Only one case (5.8%) had an IgG titre ≥800. The most common isolated valve lesion on echocardiography was severe aortic regurgitation (n=10;47.6%). Surgical replacement and repair were performed in 17(80.9%) and 4(19%) patients respectively. The one-, three- and six-month mortality was 14%, 14 % and 14% respectively. No cases of relapse were reported. Discussion B.quintana is the most prevalent cause of BCNE in our centre. The typical clinical and echocardiographic profile was consistent with the reported literature of a destructive endocarditis in a patient with a low socioeconomic background. Despite this, the surgical outcome is fair and comparable to European data. The IFA used in this study is different to the assay referenced in the ESC guideline and may explain the relatively low serology titres observed (despite PCR-proven disease). Further study with the commercial IFA is needed to determine a cut-off titre that will give a comparable sensitivity to the referenced in-house IFA.Destructive aortic valve endocarditisResultant severe aortic regurgitation

Infantile Intraparenchymal Brain Abscess due to Streptococcus pyogenes

Introduction Group A ß-hemolytic streptococcus (GABHS) are the most common bacterial cause of tonsillitis, and can cause noninvasive diseases such as pharyngitis and impetigo, as well as more severe invasive diseases. The incidence of invasive disease is 1-3/100,000 per year, and the morbidity and mortality rate is high. GABHS is rarely lead to brain abscesses. Case Report Acute phase reactants were found to be high in a 40-day-old patient who presented with fever, irritability and focal seizures. Acute phase reactants were high. Transfontanel ultrasonography showed increased thickness, echo and blood supply in the meninges. Brain magnetic resonance imaging (MRI) and diffusion MRI revealed a thick-walled abscess in the right cerebral hemisphere and parietooccipital parenchyma with diffusion restriction on diffusion-weighted images. Cerebrospinal fluid (CSF) polymerase chain reaction (PCR) test revealed S. pyogenes,and S. pyogenes grown in CSF culture. Initially, meropenem, vancomycin, metronidazole antibiotherapy was applied. Treatment was revised to cefotaxime and vancomycin after culture antibiogram and was completed for 8 weeks. Seizures regressed. Control brain imaging showed improvement. Restriction in left lower extremity movements remained. Physical therapy and rehabilitation was recommended and he was discharged. Conclusion GABHS, which is the causative agent of acute tonsillitis, rarely causes invasive disease which has high mortality and morbidity.

Investigating Transmission Patterns and Genome Sequencing of SARS-CoV-2 in University College Dublin

Abstract Background This study describes the investigation and management of multiple outbreaks of SARS-Cov2 during the second and third wave of the pandemic at University College Dublin, Ireland from September 2020 to September 2021. Methods Relevant data were gathered as part of the public health outbreak investigations led by the UCD Internal Covid Control Team (ICCT) in collaboration with the public health teams of the Health Service Executive (HSE). Results are presented for PCR (polymerase chain reaction) confirmed cases and their close contacts, reported to the UCD ICCT between September 2020 to September 2021. Results There were 214 cases notified to ICCT. Among these, 153 cases were in residence on-campus cases. 73 epidemiologically linked clusters identified, where the number of cases linked with each cluster varied between 1 to 27. Additional cases during this period had no obvious epidemiological link to the identified clusters. Of 902 close contacts with PCR test results, 31.5% (n = 284) tested positive. 77% (n = 219) self-reported with mild to moderately symptoms while 23% (n = 65) self-reported as asymptomatic. Retrospective Whole Genome Sequence (WGS) analysis was undertaken after the outbreaks had subsided. The test positive cases were grouped into 6 clusters, and it was shown that many of the apparent sporadic cases were included in these clusters. Conclusions The proportion of close contacts testing positive varied significantly throughout the pandemic, with testing policy and type of exposure having the greatest impact. Whole genome sequencing can give a better understanding of webs of transmission to complement epidemiological investigations. It is now possible to undertake sequencing in real time where it can make a contribution to outbreak control and resolution. Public Health professionals should become familiar with WGS and bioinformatics as useful tools in their armory for the control of all communicable diseases not only SARS-CoV-2. Key messages • Combined epidemiological investigations and Whole Genome Sequencing (WGS) provide crucial insights into campus COVID-19 outbreaks, guiding effective control measures. • WGS integration enhances understanding of transmission patterns, identifies clusters, and equips public health professionals with data-driven approaches for outbreak management beyond SARS-CoV-2.

Detection of Brucella militensis in the Serum of Patients Suspected of Brucellosis by the Omp31 Gene Amplification, Compared to the Serological Diagnostic Tests

Background and Objectives: Brucellosis is one of the most common infectious diseases with a global spread. It has a high prevalence in Iran. This study aims to diagnose brucellosis caused by Brucella melitensis in the serum of patients by the amplification of omp31 gene, compared to the serological tests. Methods: Blood samples were collected from 200 people suspected of brucellosis. The samples were evaluated by serological tests including Rose Bengal test, Wright test, Coombs-Wright test, gel agglutination test, and 2-mercaptoethanol (2ME) test. DNA extraction was done by the phenol/chloroform extraction method. Molecular detection with polymerase chain reaction (PCR) method was done using the specific primers of the Omp31 gene. Data were analyzed in SPSS software, version 23 using the analysis of variance. Results: Of 200 patients, 122 were female and 78 were male with a mean age of 45.17±17.43 years. The results of PCR test for the omp31 gene were positive in 14.5% of cases, which was consistent with the results of serological tests (13.8%). The sensitivity of Wright, Coombs-Wright, 2ME, and gel agglutination tests, compared to PCR, was 89.7, 75.9, 55.2 and 55.2%, respectively. Most of the affected people were housekeepers (41.5%) and urban residents (75.5%). Muscle pain (68%) and leg pain (62.3%) were the most common symptoms. Consumption of non-pasteurized dairy products was the highest risk factor (17%). Conclusion: The diagnosis of brucellosis by the omp31 gene amplification has higher sensitivity and accuracy compared to the serological tests. Considering the importance of rapid and timely diagnosis of brucellosis to control its clinical complications, the molecular diagnosis method is recommended as a diagnostic method.

Genetic profiling of virulence genes among Pseudomonas Aeruginosa isolates recovered from diseased chickens

Pseudomonas aeruginosa (P. aeruginosa) is regarded as an opportunistic organism for bird species. Also is one can be mentioned as an example of an environment-associated infection that results in severe disease in poultry farms, Indeed, we detected the incidence of P. aeruginosa in chickens and also discovered the implicated virulence genes and antibiotic-resistant mechanism. Two hundred cases were collected from broiler and layer chicken organs (heart blood, heart, and lungs) that suffered from some respiratory manifestation and septicemia in addition to yolk sacs from one-day-old chicks, which were examined bacteriologically and Serologically. The studied isolates were tested against eight antimicrobial discs. Bacterial DNA was analyzed using polymerase chain reaction (PCR) against six virulence genes (lasL, lasB toxA, exoU, exoT, and oprl genes). Eighteen isolates out of 200 (9%) of P. aeruginosa were defined. Serologically the isolates were related to more serogroup (O1,O4,O6, O11 and O12) P. aeruginosa. PCR defined the existence of P. aeruginosa by detection of the 16SRNA gene. All isolates showed 100% multidrug resistance (MDR) to more than one antimicrobial disc (sulfamethoxazole, erythromycin, tetracycline, and ampicillin). Conversely, those isolates exhibited 100% sensitivity to ciprofloxacin, and four of them showed sensitivity to norfloxacin with a percentage of (22.2%). In addition to 15 isolates showed resistance of 83.3% percent to streptomycin and nalidixic acid. By PCR test, all examined P. aeruginosa strains carried four virulence genes (lasI, toxA, exoU, and oprl). However, only three strains carried lasB, and four carried the exoT gene. In conclusion, we confirmed the presence of different factors of virulence for P. aeruginosa and MDR to more than one antimicrobial drug. So, restricting Pseudomonas infection in poultry requires strict biosecurity measures to control and eradicate the infection by P. aeruginosa

Outcomes of COVID-19 in Pregnant Women: A Retrospective Analysis of 300 Cases in Jordan

Background: The impact of COVID-19 on pregnancy remains a critical area of research, with growing evidence suggesting that maternal infection, particularly in the third trimester, may lead to significant complications Aims: The primary aim was to investigate the maternal and neonatal outcome of pregnant Jordanian women with COVID-19. The secondary aim included exploring demographics, obstetrics characteristics, and comorbidities among these women. Methods: A retrospective comprehensive review of the records of 300 cases of pregnant women with COVID-19, who were treated between November 2020 and April 2021 at Queen Alia Military Hospital (a main referral center for patients with COVID-19) in Jordan. All cases were confirmed by the rapid antigen test (RAT) + long polymerase chain reaction (PCR) test used to detect SARS-CoV-2 by amplifying viral RNA from patient samples. Women infected with COVID-19 were categorized into four groups according to the RCOG guidelines for COVID-19 infection in pregnancy: asymptomatic, mild, moderate, and severe cases. All cases were managed following the Royal College of Obstetricians and Gynecologists protocol for COVID-19 in pregnancy. Data extracted from patient’s records included demographic information, COVID-19 clinical manifestations, obstetric history, diagnostic findings, treatment plans, comorbidities, gestational age at diagnosis, treatment protocols, and maternal and neonatal outcomes. Results: The mean age was 29.7 years; 98.3% were nonsmokers; 8% had previous miscarriages, and 67.3% had the infection in the third trimester. Iron deficiency anemia affected 30.3%, while 18.3% had comorbidities, mainly hypothyroidism. Most women were asymptomatic 61.7%, but 33% had respiratory symptoms, 4.7% needed intensive care unit (ICU) admission, and 2.7% resulted in maternal deaths. First-trimester and second-trimester miscarriages were recorded in 2.67% and 3.67% of cases, respectively, while preterm labor occurred in 3.0% of pregnancies. Additionally, age and hospitalization duration had a positive correlation with the neonatal outcomes (r = 0.349, p < 0.01), (r = 0.376, p < 0.01), respectively. Furthermore, COVID-19 presentation and treatment options demonstrated a strong positive correlation (p-value <0.01). On the other hand, maternal death had a strong negative correlation with poor neonatal outcomes (r = −0.776, p < 0.01). Conclusion: The study showed that COVID-19 in pregnant women, particularly in the third trimester, is associated with significant neonatal complications, with age, hospitalization duration, and COVID-19 severity strongly impacting outcomes.

Plasmodium ovale Malaria in Travelers and Immigrants to the United States: A Case Series.

Malaria in child travelers caused by Plasmodium ovale spp. is less well characterized than malaria due to other Plasmodium species. Commonly used diagnostic tests often lack adequate sensitivity to identify P. ovale spp., and a missed diagnosis may have serious consequences. We present a case series of eight children in the United States with P. ovale malaria, all of whom had traveled to or immigrated from malaria-endemic areas. Two children developed clinical malaria, including one with severe malaria; two had isolated splenomegaly; and four were asymptomatic siblings of the children with splenomegaly. Seven of the eight children had negative blood smear readings, and the diagnosis was made by polymerase chain reaction testing. Two children had concurrent P. malariae infection despite presumptive antimalarial treatment before immigration. These findings suggest a need for reconsideration of screening and diagnostic evaluation for P. ovale malaria in high-risk groups.

Characteristics and outcomes of patients hospitalized for infection with Influenza A, SARS-CoV-2 or respiratory syncytial virus in the season 2023/2024 in a large German primary care centre

BackgroundIn addition to the persistence of SARS-CoV-2 infections, those with Influenza A/B and RSV have reappeared in 2022/23. To compare the development of prevalence, clinical outcomes and risk factors, we analysed data of the season 2023/24 from the same region/hospital as for 2022/23.MethodsPatients covering the whole age range with a positive polymerase chain reaction (PCR) test for SARS-CoV-2, Influenza A/B, RSV were included from the internal, neurological and paediatric units of the RoMed hospital Rosenheim, Germany/Bavaria, from August 1st 2023 to 29th February 2024.ResultsOf 932 patients included, 912 showed single infections with SARS-CoV-2, Influenza A or RSV (47.9% female, median age 68.0 years; 52.9% SARS-CoV-2, 23.2% Influenza A, 21.8% RSV). Co-infections (2.0%) and Influenza B (0.1%) were negligible. In patients of age ≥ 18 years (n = 628, 68.5% SARS-CoV-2, 26.0% Influenza A, 5.6% RSV), patients with Influenza A were younger compared to SARS-CoV-2 (p < 0.001), with RSV similar to SARS-CoV-2. Heart failure and asthma were the most prevalent comorbidities for RSV, immunosuppression for Influenza A. Admission to Intensive Care Unit (ICU) occurred in 111 patients (17.0% of SARS-CoV-2, 17.2% Influenza A, 28.6% RSV), and 59 patients died (8.8% SARS-CoV-2, 8.6% Influenza A, 20.0% RSV). Low-flow oxygen supplementation and non-invasive ventilation (NIV) were most frequent for RSV (68.6% and 20.0%, respectively), oxygen demand upon admission for Influenza A (39.3%), without differences in high-flow oxygen supply or length of hospital stay. Among patients aged < 18 years (n = 284, 21.4% SARS-CoV-2, 18.0% Influenza A, 57.1% RSV), 15 were admitted to ICU (4.8% SARS-CoV-2, 3.8% Influenza A, 6.0% RSV); none of them died. Oxygen supply via high-flow, low-flow or upon admission was highest for RSV (23.8%, 70.2%, 21.4%, respectively), as well as the length of hospital stay.ConclusionBetween 8/2023 to 2/2024, a large population of patients hospitalized due to respiratory tract infection, showed relative contributions of SARS-CoV-2, Influenza A or RSV similar to those in 2022/23. The findings underline that in both, adults and children, RSV posed a relatively higher clinical risk than Influenza A and SARS-CoV-2, though absolute numbers remained highest for SARS-CoV-2.

Identification of BlaTEM Gene Encode Extended Spectrum Beta Lactamase (ESBL) Producing Escherichia coli Isolated from Fresh Beef

Background: The community needs fresh beef of good quality. Dirty sanitation causes Escherichia coli to easily contaminate fresh meat and E. coli is one of the bacteria that can produce ESBL enzymes that impact human health. This study aimed to isolate Escherichia coli from fresh meat at the Surabaya Traditional Markets to identify the presence of the ESBL-encoding blaTEM gene.Methods: One hundred fifty fresh beef samples were collected from six traditional markets located in Surabaya city. Then, a polymerase chain reaction (PCR) test was performed to detect the blaTEM gene which codes for ESBL in isolates that unveiled positive results for MDR (resistant to three antibiotics) and suspected beta-lactamase enzyme (resistant to ampicillin).Results: The research showed that from 150 samples of fresh beef at the Surabaya Traditional Market, 68 Escherichia coli isolates were found (45.3%). Escherichia coli isolates were shown to be susceptible to the antibiotic’s ampicillin (73.5%), gentamicin (95%), tetracycline (7.5%), aztreonam (100%), and ciprofloxacin (95%). The highest percentage of resistance of Escherichia coli isolates to the antibiotic ampicillin (19.1%). PCR analysis of ampicillin-resistant isolates showed that the blaTEM gene encoding ESBL was present in 7 (53.8%) of the 13 Escherichia coli isolates.Conclusion: The research results show that there is a risk of ESBL transmission to the community through Escherichia coli which was identified in fresh beef at the Surabaya Traditional Market. Therefore, it is hoped that public awareness can increase regarding food safety issues which can have an impact on public health.Keywords: Escherichia coli; blaTEM; ESBL; Fresh beef; Public health

Molecular characterization and immunopathological investigation of Avian reticuloendotheliosis virus in breeder flocks in Egypt

BackgroundReticuloendotheliosis virus (REV) is an oncogenic immunosuppressive retrovirus that infects different kinds of avian species; posing significant economic losses to the poultry industry worldwide.MethodsIn Egypt, there is an unidentified disease associated with the runting-stunting syndrome with neoplasia, suspected to be REV, that has been continuously monitored in several breeder flocks. To diagnose and analyze REV by cell cultures, enzyme-linked immunosorbent assay (ELISA), histopathological investigation, the polymerase chain reaction (PCR) test, and sequencing analysis, 200 blood samples, and 50 tissue specimens were collected. The current study targets the occurrence and genetic characteristics of a viral neoplastic disease, resembling REV infection, circulating in breeder flocks from 2022 to 2023 in the Ismailia, El-Sharqia, and El-Dakahliya governorates.ResultHere, REV was isolated on chicken embryo fibroblast cell culture; exhibiting cell aggregation, rounding, and cell detachments. Collectively, only 70 serum samples were positive for anti‐REV antibodies with seroprevalence rates of 35% based on the ELISA test. The histopathological observation demonstrated lymphoreticular tumors in the liver, spleen, and other examined organs. The immunohistochemical staining method confirmed the REV-positive signals in all examined organs (liver, kidney, spleen, bursa, ovaries) except for the heart. The PCR assay of the LTR gene assessed 370 base pairs with only 5 positive samples with a percentage of 16.6%. Three positive samples were further sequenced and submitted to the Genbank under accession numbers (PP763709, PP763710, PP763711). Phylogenetic analysis of the REV-LTR gene showed that our three isolates (Sharquia-1-REV, Ismilia-2-REV, Mansoura-3-REV) are REV subtype III which predominantly circulated in breeders in Egypt. These three isolates are highest similar to American, Chinese, and Taiwanese REV reference strains, and other Egyptian strains with nucleotide identity percentages of 100%, 99%, and 99%; respectively, and on the amino acid identity level were with (99–100%), (98%, 99%), (99%, 100%); respectively.ConclusionsThis study established that REV infection was extensively distributed in the breeders and became one of the causes of the clinical outbreaks of tumors, raising awareness of REV as the causative agent of avian oncogenic disease in Egypt.

Impact of rapid blood culture identification PCR panel on optimal antibiotic use in methicillin-susceptible Staphylococcus aureus bacteremia.

In this retrospective study of 200 patients with methicillin-susceptible Staphylococcus aureus (MSSA) bacteremia, the implementation of the BioFire® blood culture identification polymerase chain reaction (PCR) panel was associated with a decreased time to optimal MSSA antibiotic therapy and shorter durations of empiric anti-MRSA antibiotic therapy and bacteremia. The findings demonstrate the significant role of rapid PCR testing and routine stewardship review in optimizing antimicrobial therapy and management of MSSA bacteremia.