Improved heat shock method for extracting total RNA from nasopharyngeal swab samples even with low viral load.

This study aimed to improve the heat shock method as a cost-effective and time-efficient for total RNA extraction. We compared the effectiveness of two total RNA extraction methods by using Real-Time PCR for nasopharynx swabs. Include: I; use of a commercial total RNA extraction kit as a standard. II; utilized a modified heat shock method (MHS). Time, centrifuge speed and duration, proteinase K, and RNA carrier were optimized. The optimized parameters included treating the sample with 5 µg/µL at 56°C for 5 minutes, heating at 95°C for 5 minutes followed by thermal shock in ice for 3 minutes, adding 4 µg/µL RNA carrier at room temperature for 3 minutes, and centrifuging at 7000 rpm for 10 minutes. This optimization demonstrated a sensitivity and specificity of 100% (CI: 95%) even in samples with low viral load. Our in-house method presents a rapid, and cost-effective alternative for total RNA extraction.

Sequence-type diversity of invasive Streptococcus pneumoniae isolates in Iran among children under 15 years of age.

Background: Infection with viruses, bacteria, or other pathogens can lead to inflammation of the meninges. Finding the pathogen and identifying the most common type is necessary for each country. Using multi-locus sequence typing (MLST), the aim of this study was to determine the genetic relationship among S. pneumoniae isolated from CSF in children with bacterial meningitis. Materials and methods: : Fourteen isolates of S. pneumoniae from CSF in children with bacterial meningitis were included in this study. The seven housekeeping genes, primer, and analysis of the sequencing used in MLST were extracted from PubMLST. Results: The sequencing analysis showed four MLST types in the studied strains. The most frequent type is ST13649 and the least frequent are ST708 and ST285. Conclusion: Finding the bacterial sequence types (ST) enables comparing the ST in different, especially neighbouring, countries.

Evaluation of Covid-19 anti-spike IgG antibody five months after the second Covid-19 vaccination.

Background: Studies in different communities have shown significant differences in IgG antibody titers in the time period after the first and second doses of the vaccines. This study aimed to serologically evaluate the IgG anti-spike antibody titer five months after injection of the second COVID-19 vaccine in healthcare workers. Materials and method: This study was performed in healthcare personnel for whom five months had passed since their second anti-Covid-19 vaccination. The level of IgG antibody against SARS-CoV-2 spike protein was measured by ELISA. Healthcare workers in Mofid Children's hospital received three brands of vaccines: Sputnik V, Sinopharm, and AstraZeneca. Results: The mean titer of anti-spike IgG was 4.3±2.29 units. The percentage of positive cases of the antibody was estimated to be 96.4%. The titer of anti-spike IgG antibody was dependent on both the occupational area and a positive history of Covid-19 disease. Conclusion: About 96.4% of the staff vaccinated against Covid-19 had a high titer of anti-spike IgG antibody even five months after inoculation of the second dose. The field of occupational can affect the level of antibody present.

Seroepidemiological and Molecular Survey for the Detection of SARS-CoV-2 Infection among Children in Iran, September 2020 to June 2021: 1-Year Cross-Sectional Study.

A population-based seroepidemiological and molecular survey on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was performed to detect induced antibodies to prior exposure and active infection of children aged 14 years or less in Tehran between 19 September 2020 and 21 June 2021. Moreover, correlations between the children's demographic data and coronavirus disease 2019 (COVID-19) symptoms with the infection status were investigated. Out of 1517 participants, cardinal symptoms of COVID-19 (fever > 38 °C and/or cough and/or diarrhea) were detected in 18%, and serological history of SARS-CoV-2 infection and polymerase chain reaction (PCR) positivity were confirmed in 33.2% and 10.7% of the weighted population, respectively. The prevalence of SARS-CoV-2 infection was significantly higher among 10-14-year-old children. Active infection was significantly higher in symptomatic children and during autumn 2020 and spring 2021. The quantitative reverse transcription real-time PCR (RT-qPCR) positivity was significantly higher among families with a lower socioeconomic status, whereas no association between RT-qPCR or seropositivity was determined with household size, underlying diseases, or gender. In conclusion, high SARS-CoV-2 infection prevalence and seroprevalence were detected in children in Tehran in different seasons. Infection prevalence was significantly higher in older children and in those with a positive history of close contact with infected cases and/or lower socioeconomic status.

Antibiotic Susceptibility Patterns for Carbapenem-Resistant Enterobacteriaceae.

Carbapenem is a broad-spectrum beta-lactam antibiotic considered the last choice for the treatment of antibiotic-resistant Gram-negative bacteria. Thus, the increasing rate of carbapenem resistance (CR) in Enterobacteriaceae is an urgent public health threat. This study aimed to evaluate the antibiotic susceptibility pattern of carbapenem-resistant Enterobacteriaceae (CRE) to new and old antibiotics. In this study, Klebsiella pneumoniae, E. coli, and Enterobacter spp. were collected from 10 hospitals in Iran for one year. CRE is recognized by resistance to meropenem and/or imipenem disk after identification of the collected bacteria. Antibiotic susceptibility of CRE against fosfomycin, rifampin, metronidazole, tigecycline, and aztreonam was detected by disk diffusion method and colistin by MIC. In this study, 1222 E. coli, 696 K. pneumoniae, and 621 Enterobacter spp. were collected from 10 hospitals in Iran in one year. Fifty-four E. coli (4.4%), 84 K. pneumoniae (12%), and 51 Enterobacter spp. (8.2%) were CRE. All CRE strains were resistant to metronidazole and rifampicin. Tigecycline has the highest sensitivity on CRE and levofloxacin for Enterobacter spp. Tigecycline showed an acceptable effectiveness rate of sensitivity on the CRE strain. Therefore, we suggest that clinicians consider this valuable antibiotic to treat CRE.

Assessment of early and post COVID-19 vaccination antibody response in healthcare workers: a multicentre cross-sectional study on inactivated, mRNA and vector-based vaccines.

In this multicentre study, we compared the status of antibody production in healthcare personnel (HCP) before and after vaccination using different brands of COVID-19 vaccines between March 2021 and September 2021. Out of a total of 962 HCP enrolled in our study, the antibody against the S1 domain of SARS-CoV-2 was detected in 48.3%, 95.5% and 96.2% of them before, after the first and the second doses of the vaccines, respectively. Our results showed post-vaccination infection in 3.7% and 5.9% of the individuals after the first and second doses of vaccines, respectively. The infection was significantly lower in HCP who presented higher antibody titres before the vaccination. Although types of vaccines did not show a significant difference in the infection rate, a lower infection rate was recorded for AstraZeneca after the second vaccination course. This rate was equal among individuals receiving a second dose of Sinopharm and Sputnik. Vaccine-related side effects were more frequent among AstraZeneca recipients after the first dose and among Sputnik recipients after the second dose. In conclusion, our results showed diversity among different brands of COVID-19 vaccines; however, it seems that two doses of the vaccines could induce an antibody response in most of HCP. The induced immunity could persist for 3-5 months after the second vaccination course.