Coronavirus disease 2019 (COVID-19): A New Pandemic and its Challenges

Emerging of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a worldwide outbreak and a major public health problem. The present review was conducted to provide brief information about the origin, symptoms, transmission, pathogenesis, diagnosis, and treatment of the virus. A search was performed in the databases of PubMed, Scopus, Science Direct, and Google scholar with English keywords including 2019-nCoV, Coronavirus disease 2019 (COVID-19), SARS-CoV-2, and novel coronavirus from December 2019 to 15 March 2020, and the search results were evaluated. Selected studies have shown that the virus may have originated from the bat. It has also been shown that the virus receptor is angiotensin-converting enzyme 2 (ACE2) which is also the SARS virus receptor and is expressed in most human tissues. The most common way of virus transmission was suggested through respiratory droplets and close contact. It is also transmitted by asymptomatic patients, but vertical transmission from mother to fetus has not been confirmed. Real-time reverse transcriptase (RT)-PCR is the gold standard for SARS-CoV-2 detection, but chest computed tomography (CT) can be more sensitive to detect positive cases. Since no effective vaccine or drug for prevention and treatment of this disease has not yet been identified and also because of the high incubation and infection period, easy transmission, and the lack of complete recognition of the characteristics and stability in different environments, the best way to control of COVID-19 is to prevent the spread of the infection in different ways and take seriously personal and public hygiene

Genetic Characterization of blaSHV/VEB/PER Genes in ESBL-producing MDR Klebsiella Pneumonia Strains Isolated from Patients in Isfahan, Iran

This study was conducted to detect three genetical variants of Extended-Spectrum Beta-Lactamase (ESBL, in which142 Klebsiella pneumoniae (K.pneumoniae) isolates were collected from sections of a teaching hospital in Isfahan and were detected using standard IMVIC biochemical tests and urease. These were confirmed by identification of the ureD gene. Antimicrobial susceptibility testing was performed using the standard Kirby-Bauer disk-diffusion method on Mueller-Hinton agar (Merck,Germany) .The performance and interpretation were based on the guidelines from the Clinical Laboratory Standards Institute (CLSI, 2013). Screening and phenotypic identification of ESBLs isolates were performed by DDST. The presence of genes responsible for ESBL resistance, such as SHV, PER and VEB type ESBL genes was identified by PCR and indicator isolates sequencing performed by Macrogen (Seoul, Korea). The nucleotide sequences were analysed using the BLAST program (http://www.ncbi.nlm.nih.gov/BLAST), the Lahey database, and CromasPro-2 and Mega-4 software to determine the subvarients of the three variants of ESBL (SHV, PER, VEB). These were compared with blaSHV-11 gene from K. pneumonia (accession.no.X98101), blaSHV-5 gene from K. pneumonia (accession no. X55640), blaPER-1 gene from P.aeruginosa transposed on Tn2345 (accession no AY866517.2) and blaVEB-1 gene from K. pneumonia (accession no. AF010416). A total of 120 isolates (84%) were recognized as MDR. The highest rate of resistance was recorded for piperacillin (80%), ceftazidime (76%), and cefotaxime (73%) and the lowest rate was for ertapenem (47.3%), meropenem (50.8%), and imipenem (58.7%) following detection of ESBL isolates of K.pneumoniae (101 isolates; 71%).The ward and the clinical specimen with the most prevalence were ICU with 55(38.7%) and urine with 61(42.9%). The lowest prevalence was related to the neurosurgery ward with 8(5.6%) samples and the clinical specimen with the lowest prevalence was cerebrospinal fluid (CSF) with 2 (1.4%) samples. PCR detection in ESBL-producing K. pneumoniae showed that, of the clinical isolates, 42.2% contained blaSHV (42/101), 2.9% contained blaVEB (3/101) and2% contained blaPER (2/101). Sequencing of 10 selected PCR products of SHV genes showed that 7/10isolates were similar to the strain SHV-11 and 3/7 isolates were similar to the strain SHV-5. The sequencing of two PCR products of the PER genes showed they were similar to the strainPER-1.Sequencing of three PCR products of the VEB genes showed they were similar to the strain VEB-1. The overall prevalence of ESBL-producers was found to vary greatly in different geographical areas; this may be the result of differences in the type and amount of antibiotics consumed and differences in the time of collection of isolates. The present study reflects anincrease in the prevalence of ESBL-producers in Iran. The most common ESBL type found in this study was SHVand that VEB and PER types were rare. In addition, sequence analysis results of our study show the rate of SHV-11, PER-1 and VEB-1was maximum

Genetic Characterization of blaSHV/VEB/PER Genes in ESBL-producing MDR Klebsiella Pneumonia Strains Isolated from Patients in Isfahan, Iran

This study was conducted to detect three genetical variants of Extended-Spectrum Beta-Lactamase (ESBL, in which142 Klebsiella pneumoniae (K.pneumoniae) isolates were collected from sections of a teaching hospital in Isfahan and were detected using standard IMVIC biochemical tests and urease. These were confirmed by identification of the ureD gene. Antimicrobial susceptibility testing was performed using the standard Kirby-Bauer disk-diffusion method on Mueller-Hinton agar (Merck,Germany) .The performance and interpretation were based on the guidelines from the Clinical Laboratory Standards Institute (CLSI, 2013). Screening and phenotypic identification of ESBLs isolates were performed by DDST. The presence of genes responsible for ESBL resistance, such as SHV, PER and VEB type ESBL genes was identified by PCR and indicator isolates sequencing performed by Macrogen (Seoul, Korea). The nucleotide sequences were analysed using the BLAST program (http://www.ncbi.nlm.nih.gov/BLAST), the Lahey database, and CromasPro-2 and Mega-4 software to determine the subvarients of the three variants of ESBL (SHV, PER, VEB). These were compared with blaSHV-11 gene from K. pneumonia (accession.no.X98101), blaSHV-5 gene from K. pneumonia (accession no. X55640), blaPER-1 gene from P.aeruginosa transposed on Tn2345 (accession no AY866517.2) and blaVEB-1 gene from K. pneumonia (accession no. AF010416). A total of 120 isolates (84%) were recognized as MDR. The highest rate of resistance was recorded for piperacillin (80%), ceftazidime (76%), and cefotaxime (73%) and the lowest rate was for ertapenem (47.3%), meropenem (50.8%), and imipenem (58.7%) following detection of ESBL isolates of K.pneumoniae (101 isolates; 71%).The ward and the clinical specimen with the most prevalence were ICU with 55(38.7%) and urine with 61(42.9%). The lowest prevalence was related to the neurosurgery ward with 8(5.6%) samples and the clinical specimen with the lowest prevalence was cerebrospinal fluid (CSF) with 2 (1.4%) samples. PCR detection in ESBL-producing K. pneumoniae showed that, of the clinical isolates, 42.2% contained blaSHV (42/101), 2.9% contained blaVEB (3/101) and2% contained blaPER (2/101). Sequencing of 10 selected PCR products of SHV genes showed that 7/10isolates were similar to the strain SHV-11 and 3/7 isolates were similar to the strain SHV-5. The sequencing of two PCR products of the PER genes showed they were similar to the strainPER-1.Sequencing of three PCR products of the VEB genes showed they were similar to the strain VEB-1. The overall prevalence of ESBL-producers was found to vary greatly in different geographical areas; this may be the result of differences in the type and amount of antibiotics consumed and differences in the time of collection of isolates. The present study reflects anincrease in the prevalence of ESBL-producers in Iran. The most common ESBL type found in this study was SHVand that VEB and PER types were rare. In addition, sequence analysis results of our study show the rate of SHV-11, PER-1 and VEB-1was maximum

Application of Epstein-Barr Virus for Optimization of Immortalized B-lymphocyte Production as a Positive Control in Genetic Studies.

BACKGROUND Infection of B-cells with Epstein-Barr virus (EBV) leads to more and subsequent immortalization. This is considered as the method of choice for generating lymphoblastoid cell lines (LCLs). Producing LCLs, although very useful but is very time consuming and troublesome, drives the requirement for quicker and more reliable methods for EBV-driven B-cell transformation. MATERIALS AND METHODS After successfully production of LCLs, different parameters including temperature, serum concentration, type of culture medium, and CO2 concentration were evaluated on EBV-transformed B-cells. In this study, we were able to produce LCLs and optimize condition. RESULTS The best condition for generating LCLs was 37°C, 5% CO2, 20% fasting blood sugar, and RPMI 1640. The study results were to establish a reliable method for producing LCLs that can be used to produce immortalized B-cells from almost any sources. CONCLUSION This can help with tumorgenecity studies, as well as producing control material for rare genetic disorders and so on. The aim of this study was to determine optimized condition for reliable and reproducible LCLs from different sources

Antimicrobial Resistance Pattern and Spectrum of Multiple-drug-resistant Enterobacteriaceae in Iranian Hospitalized Patients with Cancer

Background: Nosocomial infections are one of the most leading causes of morbidity and mortality in patients with cancer. The emergence of multiple-drug-resistant (MDR) strains of Gram-negative bacteria causing nosocomial infection has become a serious concern in cancer patients. Therefore, the present study aimed to determine the spectrum and antibiotic resistance pattern of Gram-negative bacteria related nosocomial infections among Iranian cancer patients. Materials and Methods: This descriptive cross-sectional study was conducted during the 6 months from December 2015 to May 2016 in two tertiary care centers located in Isfahan and Arak Province. Gram-negative bacteria obtained from different clinical specimens from hospitalized patients with cancer and were identified using standard microbiological methods. Antibiotic susceptibility pattern was determined by the disk diffusion method according to the Clinical and Laboratory Standards Institute (CLSI) recommendation. Results: Of totally 259 culture positive cases, Escherichia coli showed the highest isolation rate (60.6%) followed by Klebsiella pneumoniae (26.6%) and Proteus spp (11.2%). The rate of MDR isolates were 91.5% (237/259). Overall, the most frequent source of bacterial isolation was urinary tract infection (65.6%) followed by skin and soft-tissue infection (23.6%). The antibiotic susceptibility results showed meropenem (MEN) and ceftazidime as the most effective antibiotics for E. coli, K. pneumoniae, and Proteus spp. isolates. Moreover, MEN was the most effective antibiotic against MDR isolates. Conclusion: The study findings showed a significant distribution of MDR Gram-negative bacteria which may increase the burden of healthcare-associated infections in cancer patients. Although, carbapenem can be considered as effective agents toward MDR strains for empirical antibiotic therapy in our region

Urinary BK virus excretion in children newly diagnosed with acute lymphoblastic leukemia

Background: Determining the risk factors in developing or increasing the relapses of acute lymphoblastic leukemia (ALL) may help health and preventive systems to launch new programs. Up to 90% of normal population changes to seropositive for BK virus by the age of 10 years. Whether this oncogenic virus is responsible for evolving ALL is unclear. In this study, we evaluated the excretion of urinary BK virus in newly diagnosed children with ALL compared with normal population. Methods: This case-control study was carried out on 62 participants (32 ALL patients and 32 normal subjects), aged 1-18 years, in Saint Al-Zahra and Sayyed-Al-Shohada University Hospitals, Isfahan, Iran. A polymerase chain reaction (PCR) method was used to detect the BK virus in specimens. PCR amplification was performed using specific primers of PEP-1 (5′-AGTCTTTAGGGTCTTCTACC-3′) and PEP-2 (5′-GGTGCCAACCTATGGAACAG-3′). Results: Thirty-five out of 62 participants (54.8%) were males and the remaining were females. The mean duration of disease was 9.6 ± 9.69 months. Central nervous system (CNS) relapse was seen in 29% of the patients. Positive PCR for urine BK virus was seen in three children with ALL (9.7%). No positive result for urine BKV was achieved in the control group. However, Fisher′s exact test did not show any significant difference between the two groups (P > 0.05). In addition, there was no significant correlation between BKV positivity and frequency of relapses. Conclusion: To demonstrate the role of BK virus in inducing ALL or increasing the number of relapses, prospective studies on larger scale of population and evaluating both serum and urine for BK virus are recommended

Genetic Diversity of Drug-resistant Mycobacterium tuberculosis Isolates in Isfahan Province of Iran

Background: Increasing drug resistance is an important factor in the complexity of tuberculosis (TB) control. The identification of disease transmission type, recurrence of a previous infection, or new transmission of the disease is the key factor in the control of TB. In this study, we aimed to identify the genetic diversity of drug-resistant Mycobacterium tuberculosis isolates in Isfahan province of Iran through the mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing method based on 24 loci. Materials and Methods: Of 300 isolates obtained from a variety of clinical specimens, 18 drug-resistance M. tuberculosis clinical isolates (resistant to a single drug to more than one drug) were collected between 2013 and 2015 from regional TB reference laboratory in Isfahan. All drug-resistance M. tuberculosis isolates were typed by 24-locus MIRU-VNTR typing. Results: The highest percentage of isolates, 38.8%, belonged to the East-Asian lineage (lineage 2), while the lineages Indo-Oceanic (lineage 1), East-African–Indian (lineage 3), and Euro-American (lineage 4) represented 5.5%, 22.2%, and 33.3%, respectively. Among the 33.3% (6/18) Euro-American strains, the Latin American– Mediterranean and Ural sub-lineage were 22.2% (4/18) and 11.1% (2/18), respectively. Conclusion: The results of this study show that the lineages of drug-resistant M. tuberculosis isolates in Isfahan province of Iran are similar to those reported in the Eastern Mediterranean region (indicative of the epidemiological relationship between the countries in the region). Continued molecular monitoring is important as it has been proposed that the genetics and evolutionary backgrounds of drug-resistant M. tuberculosis strains may have an impact on the transmissibility profile

Evaluation of the effect of Pulicaria gnaphalodes and Perovskia abrotanoides essential oil extracts against Mycobacterium tuberculosis strains

Background: Mycobacterium tuberculosis (MTB) is the causative agent of tuberculosis (TB), which remains one of the major public health problems in the world. The increasing incidence of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) worldwide highlights the urgent need to search for alternative antimycobacterial agents. More and more people in developing countries utilize traditional medicine for their major primary health care needs. It has been determined that the medicinal plants Pulicaria gnaphalodes and Perovskia abrotanoides possess strong antibacterial effect. Materials and Methods: In this study, the antimycobacterial effects of P. gnaphalodes and P. abrotanoides essential oil on MTB were examined. Essential oil was prepared from P. gnaphalodes aerial parts and P. abrotanoides flower. The effects of six different concentrations (20 μg/ml, 40 μg/ml, 80 μg/ml, 160 μg/ml, 320 μg/ml, and 640 μg/ml) were examined against sensitive isolates of MTB and MTB H37Rv (ATCC 27294). Results: The results showed that P. gnaphalodes and P. abrotanoides essential oil extracts have strong inhibitory effects on MTB. This activity for P. gnaphalodes was observed from very low (4%) to good (70.9%) effect; meanwhile, this activity for P. abrotanoides was observed from very low (4%) to strong (86%) effect. Conclusion: The mean of inhibition percentage for P. gnaphalodes and P. abrotanoides in 640 μg/ml was 58.1% and 76.2%, respectively. So, P. abrotanoides plant is more effective against MTB than P. gnaphalodes. Identification of the effective fraction against MTB is a further step to be studied