A rapid influenza diagnostic test based on detection of viral neuraminidase activity

Current methods used for diagnosis of acute infection of pathogens rely on detection of nucleic acids, antigens, or certain classes of antibodies such as IgM. Here we report a virus enzyme assay as an alternative to these methods for detection of acute viral infection. In this method, we used a luciferin derivative as the substrate for detection of the enzyme activity of influenza viral neuraminidase as a means for diagnosis of influenza. The resulting commercial test, the qFLU Dx Test, uses a different supply chain that does not compete with those for the current tests. The assay reagents were formulated as a master mix that accommodated both the neuraminidase and luciferase reactions, thereby enabling rapid and prolonged production of stable light signal in the presence of influenza virus in the sample. The assay was evaluated using depository throat swab specimens. As expected, the assay exhibited similar detection rates for all influenza types and subtypes except for A(H7N9), which exhibited lower detection rate due to lower viral titer in the specimens. When throat swab specimens were diluted with the sample buffer of the test kit and tested with the qFLU Dx Test. The sensitivity and specificity were 82.41% (95% confidence interval: 79.66–85.84%) and 95.39% (95% confidence interval: 94.32–96.46%), respectively, for these diluted specimens in comparison to a real-time polymerase chain reaction assay. The uniqueness of the qFLU Dx Test as an enzymatic assay makes it highly complementary with currently available methods.

High Dilutions of a Drug of Covid-19 Origin Show Difference in Ranks

High dilutions (HDs) of drugs, used in Homeopathy, are prepared in aqueous EtOH (ethanol) through serial dilution accompanying mechanical agitation or succussion, and are called potencies. The potencies from the rank 12 onwards are too dilute to contain any original drug molecules. Do the potency ranks show any difference from each other? Do serial dilution and succussion contribute to the difference in potency ranks? This study aims to address these two questions. The throat swab of a Covid-19 patient was preserved and diluted with aqueous EtOH 90% to prepare the mother tincture (MT) and five different potencies of Covid named Covidinum. These potencies and their solvent media were analysed by electronic and vibrational spectroscopy. Charge transfer (CT) and proton transfer interactions occur during preparation of the potencies. The FT-IR spectra of all the test samples after normalization show difference from each other with respect to O-H stretching and bending (v2) bands. Serial dilution and succussion contribute to the observed difference in ranks and CT interactions.

Determination of antimicrobial activity of traditional spices extracts against clinical isolates in Dhaka city

Spices are dried aromatic substances derived from plants and have demonstrated antimicrobial activity against pathogenic microorganisms. In the present study, a total of six spices turmeric (Curcuma longa), garlic (Allium sativum), black pepper (Piper nigrum), ginger (Zingiber officinale), clove (Syzygium aromaticum) and cinnamon (Cinnamomum verum) were evaluated for their antibacterial activity. Antibacterial activities of ethanol, propanol and water extracts were determined by agar well diffusion assay against previously isolated Klebsiella pneumoniae, Staphylococcus aureus from urine samples and Citrobacter spp. from throat swab samples. All spices examined in this study showed antibacterial activity in different types of extraction methods. Water, ethanol and propanol extracts exhibited anti-bacterial activity measured as zone of inhibition between 6 mm and 25 mm in diameter. Water extract of garlic (Allium sativum) showed the highest antibacterial activity (25mm) against clinical isolates of Staphylococcus aureus and Citrobacter spp. Water extract of other species showed the lowest antibacterial activity (6 mm) against throat swab isolates Staphylococcus aureus and Citrobacter spp. Spices which showed signifiant antimicrobial activities can be further studied for the isolation of active ingredients and development of novel drugs. Stamford Journal of Microbiology, Vol.11 (1) 2021: 17-19

Modern understanding about the role of group A β -hemolytic streptococcus in acute tonsillitis

Objective: assess the modern value of group А β-hemolytic streptococcus in patients with acute tonsillitis and the effectiveness of the rapid streptococcal antigen detection method.Materials and methods: microbial landscape assessment of acute tonsillitis was based on retrospective analysis of 902 bacterial culture results of a throat swab of patients with syndromes of acute tonsillitis treated in the Infectious Diseases Clinic of the Military Medical Academy named after S.M. Kirov during the period of 2019-2020. The effectiveness of the rapid streptococcal antigen detection method in the oropharynx was determined by a prospective study involving 35 patients with acute tonsillitis.Results: in the study, we have found that bacterial culture results of a throat swab, the following were more common: Nesseria species (39 %), Streptococcus viridans (23 %), and Staphylococcus aureus (17 %). The frequency of detection of β-hemolytic streptococcus was 1 %. The rapid diagnostic system «Streptatest» in patients with acute tonsillitis has demonstrated efficiency, under which that sensitivity of test was 80 %, specificity – 90 %, positive predictive value – 57,14 %, negative predictive value – 96,43 %.Conclusions: the frequency of group A β-hemolytic streptococcus in patients with lesion of lymphoid tissues of the oropharynx has declined significantly nowadays. The rapid diagnostic system «Streptatest» is a highly effective medical product that can be used in both hospital and pre-hospital stage.

Preceding group A streptococcus skin and throat infections are individually associated with acute rheumatic fever: evidence from New Zealand

IntroductionAcute rheumatic fever (ARF) is usually considered a consequence of group A streptococcus (GAS) pharyngitis, with GAS skin infections not considered a major trigger. The aim was to quantify the risk of ARF following a GAS-positive skin or throat swab.MethodsThis retrospective analysis used pre-existing administrative data. Throat and skin swab data (1 866 981 swabs) from the Auckland region, New Zealand and antibiotic dispensing data were used (2010–2017). Incident ARF cases were identified using hospitalisation data (2010–2018). The risk ratio (RR) of ARF following swab collection was estimated across selected features and timeframes. Antibiotic dispensing data were linked to investigate whether this altered ARF risk following GAS detection.ResultsARF risk increased following GAS detection in a throat or skin swab. Māori and Pacific Peoples had the highest ARF risk 8–90 days following a GAS-positive throat or skin swab, compared with a GAS-negative swab. During this period, the RR for Māori and Pacific Peoples following a GAS-positive throat swab was 4.8 (95% CI 3.6 to 6.4) and following a GAS-positive skin swab, the RR was 5.1 (95% CI 1.8 to 15.0). Antibiotic dispensing was not associated with a reduction in ARF risk following GAS detection in a throat swab (antibiotics not dispensed (RR: 4.1, 95% CI 2.7 to 6.2), antibiotics dispensed (RR: 4.3, 95% CI 2.5 to 7.4) or in a skin swab (antibiotics not dispensed (RR: 3.5, 95% CI 0.9 to 13.9), antibiotics dispensed (RR: 2.0, 95% CI 0.3 to 12.1).ConclusionsA GAS-positive throat or skin swab is strongly associated with subsequent ARF, particularly for Māori and Pacific Peoples. This study provides the first population-level evidence that GAS skin infection can trigger ARF.

Evaluation of Rapid Antigen Detection Test for Group A Streptococci Pharyngitis among Children in an Out-Patient Clinic in Malaysia

One of the most common conditions encountered in the out-patient setting is acute pharyngitis. Group A Streptococcus (GAS) accounts for 15%-30% of cases of sore throat particularly in children under 15 years old. Rapid antigen testing (RADT) is an alternative diagnostic method to detect GAS pharyngitis. This study was done to evaluate the agreement between RADT whereby BIONEXIA® Strep A Plus (BioMérieux, France) kit was used and throat culture in the diagnosis of GAS pharyngitis in children presented with a sore throat. One hundred and ten children from a primary health care clinic with sore throat were included in this study. All children were evaluated based on McIsaac scoring and throat swab samples were taken for both throat culture and RADT testing. The prevalence of GAS pharyngitis by RADT in this study was 7.3% over one year. A higher incidence of GAS pharyngitis was noted in the school-aged children than the preschool-age children. There was no correlation between cough, lymph node enlargement, and tonsillar enlargement in predicting GAS pharyngitis. The sensitivity and specificity of RADT were 100% and 98%, respectively, when taking throat culture as a gold standard. A good agreement between RADT and throat culture was achieved (k=0.848). McIsaac scoring was noted to have good predictability for GAS pharyngitis with AUC=0.82. In conclusion, the rapid streptococcal antigen detection test showed excellent sensitivity and specificity and detecting GAS from the throat swab samples. Thus, it can be used to aid in the diagnosis of group A Streptococcal pharyngitis and could reduce the overuse of antibiotics. McIsaac score has also proven to be useful as a screening tool for bacterial pharyngitis.

Case Report: Genomic Characteristics of the First Known Case of SARS-CoV-2 Imported From Spain to Sichuan, China

The pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been basically under control in China since March 2020, but the import of domestic SARS-CoV-2 has begun to increase. This study reported the first case of asymptomatic SARS-CoV-2 infection imported from Spain into Sichuan Province, China, on March 11, 2020. The infected male had a body temperature of 37.5°C, normal blood oxygen saturation levels, and a computed tomography (CT) examination showed that his lungs had no shadows. However, a throat swab from the subject tested positive for SARS-CoV-2 using qPCR assay. In this study, we conducted transcriptome sequencing on respiratory throat swabs from the subject and found that the dominant SARS-CoV-2 sequence (Gene Bank ID: MW301121) was a spike protein D614G mutant strain, which is currently popular throughout world. We downloaded and analyzed SARS-CoV-2 sequences collected from cases in China and Spain for comparison and tracing purposes. After March 11, 2020, the Chinese domestic clade was naturally divided into the imported SARS-CoV-2 D614G mutant strain and evolutionarily-related similar sequences and that of sequences collected in the original Wuhan area. The sequence reported in this study was located on a small branch, far from the evolution of Wuhan sequences. As expected, the identified sequence was closely related to the evolution of the SARS-CoV-2 D614G mutant strain circulating in Spain.

Reduced seroconversion in children compared to adults with mild COVID-19

Importance: The immune response in children with SARS-CoV-2 infection is not well understood. Objective: To compare seroconversion in children and adults with non-hospitalized (mild) SARS-CoV-2 infection and to understand the factors that influence this. Design: Participants were part of a household cohort study of SARS-CoV-2 infection. Weekly nasopharyngeal/throat swabs and blood samples were collected during the acute and convalescent period following PCR diagnosis for analysis. Setting: Participants were recruited at the Royal Childrens Hospital, Melbourne, Australia between May and October 2020. Participants: Those who had a SARS-CoV-2 PCR-positive nasal/throat swab. Main outcomes and measures: SARS-CoV-2 antibody and cellular responses in children and adults. Seroconversion was defined by seropositivity in all three serological assays. Results: Among 108 SARS-CoV-2 PCR-positive participants, 57 were children (median age: 4, IQR 2-10) and 51 were adults (median age: 37, IQR 34-45). Using three established serological assays, a lower proportion of children seroconverted compared with adults [20/54 69 (37.0%) vs 32/42 (76.2%); (p<0.001)]. This was not related to viral load, which was similar in children and adults [mean Ct 28.58 (SD: 6.83) vs 24.14 (SD: 8.47)]. Age and sex also did not influence seroconversion or the magnitude of antibody response within children or adults. Notably, in adults (but not children) symptomatic adults had three-fold higher antibody levels than asymptomatic adults (median 227.5 IU/mL, IQR 133.7-521.6 vs median 75.3 IU/mL, IQR 36.9-113.6). Evidence of cellular immunity was observed in adults who seroconverted but not in children who seroconverted. Conclusion and Relevance: In this non-hospitalized cohort with mild COVID-19, children were less likely to seroconvert than adults despite similar viral loads. This has implications for future protection following COVID-19 infection in children and for interpretation of serosurveys that involve children. Further research to understand why children are less likely to seroconvert and develop symptoms following SARS-CoV-2 infection, and comparison with vaccine responses may be of clinical and scientific importance.

A STUDY OF NEONATAL OUTCOMES IN BABIES BORN TO COVID- 19 POSITIVE MOTHERS

MATERIAL & METHOD: This retrospective study was conducted during the period of June-August 2020 for 3months in newborns born to COVID-19 positive mothers at YMCH after obtaining ethical clearance. All newborns born to mothers with COVID-19 positive status during any trimester of pregnancy, irrespective of neonatal COVID-19 RT-PCR test, and delivered at YMCH during the study period were included. Relevant history and complete systemic examination ndings of the neonate were noted down from the case records and laboratory results of RT-PCR test on nasopharyngeal and throat swab if done was noted down which was done at 24-48hours of birth.RESULT:Total of 25newborns born to COVID-19 positive mothers were included in the study. Among the neonates born, 64% were female and 36% were male. 76% of the pregnant mothers were asymptomatic at the time of detection of COVID-19 infection. The neonatal outcome found to have only 1 newborn with positive COVID-19 infection compared to 24 healthy newborns. CONCLUSION: Signicant measures for preventing neonatal SARS-CoV-2 infection are to prevent maternal illness and to decrease the potential of neonatal viral exposure.