Background Antibiotic resistance is currently the most serious global threat to the effective treatment of bacterial infections. Antibiotic resistance has been established to adversely affect both clinical and therapeutic outcomes, with consequences ranging from treatment failures and the need for expensive and safer alternative drugs to the cost of higher rates of morbidity and mortality, longer hospitalization, and high‐healthcare costs. The search for new antibiotics and other antimicrobials continues to be a pressing need in humanity's battle against bacterial infections. Antibiotic resistance appears inevitable, and there is a continuous lack of interest in investing in new antibiotic research by pharmaceutical industries. This review summarized some new strategies for tackling antibiotic resistance in bacteria. Methods To provide an overview of the recent research, we look at some new strategies for preventing resistance and/or reviving bacteria's susceptibility to already existing antibiotics. Results Substantial pieces of evidence suggest that antimicrobials interact with host immunity, leading to potent indirect effects that improve antibacterial activities and may result in more swift and complete bactericidal effects. A new class of antibiotics referred to as immuno‐antibiotics and the targeting of some biochemical resistance pathway components including inhibition of SOS response and hydrogen sulfide as biochemical underlying networks of bacteria can be considered as new emerging strategies to combat antibiotic resistance in bacteria. Conclusion This review highlighted and discussed immuno‐antibiotics and inhibition of SOS response and hydrogen sulfide as biochemical underlying networks of bacteria as new weapons against antibiotic resistance in bacteria.
There is still no agreement on the gold standard technique for diagnosing of H. Pylori in Iraq, as well as a paucity of data on the validity of different diagnostic techniques. This study aimed to investigate the prevalence of this bacterium with different methods and compare them to the quantitative polymerase chain reaction (qPCR) as a golden standard technique among Iraqi patients. In total, 115 Iraqi patients strongly suspected of H. pylori infection were enrolled in the current study. Invasive techniques including rapid urease testing (RUT) and gastric tissue culture in addition to non-invasive techniques including 14C-Urea breath test (14C-UBT), stool antigen test (SAT), CagA-IgG serology, and qPCR were performed to confirm the H. pylori infection. The qPCR was used as the gold standard to estimate the sensitivity, specificity, positive and negative predictive values for the studied diagnostic tests. Overall, the prevalence rate of H. pylori in Iraqi patients was ranged from 47.8 to 70.4% using different methods. The positive results for each test were as follows: qPCR 81, (70.4%) UBT 79 (68.7%), SAT 77 (67%), RUT 76 (66.1%), Cag-IgG 61 (53%), and culture 55 (47.8%). The 14C-UBT showed the highest overall performance with 97.5% sensitivity, 97% specificity, and total accuracy of 97.3% followed by SAT, RUT, Cag-IgG, and culture method. Based on the accuracy of the studied methods for H. pylori detection, they can be arranged in order as follows: qPCR > UBT > SAT > RUT> CagA IgG > culture. Since a universal gold standard assay for the diagnosis of H. pylori has not yet been established in Iraq, the UBT may be recommended as first choice due to its higher performance compared to other methods.
Objective: Multi-drug resistance Gram-negative bacteria possessing Extended-Spectrum β-Lactamase (ESBL) genes are of concern because of their resistance to third-generation cephalosporins. This study aims to investigate the molecular basis of resistance to modern β-lactams by ESBLs encoded by the blaVEB gene and the gene’s role in resistance. Also, gene sequencing was used to compare genetic similarities with global isolates using phylogenetic and cluster analyses. Methods: Between March and July 2018, a total of 100 Iraqi clinical isolates were examined, in this cross-sectional study, to determine their ESBL status using the double-disc synergy technique. Polymerase Chain Reactions (PCRs) were performed on extracted blaVEB genes and sequencing of the target PCR products was performed. All blaVEB sequences were compared with the available sequence data, using BLAST searches against the GenBank database. Results: A total of 35 isolates, comprising 5 Escherichia coli, 18 Klebsiella pneumoniae, and 12 Pseudomonas aeruginosa isolates were confirmed to possess ESBLs; the blaVEB gene was detected in one isolate of each species. The sequencing of these genes revealed 99% similarity with the global standard genes deposited in GenBank. Conclusion: The blaVEB gene plays an essential role in the resistance of ESBL-producing isolates to new β-lactams. Further, the sequencing and phylogenetic analyses of the genes from the P. aeruginosa, K. pneumonia, and E. coli isolates revealed 99% similarity with the GenBank global standard genes.
Purpose This study aimed to evaluate the presence of CRISPR-Cas system genes and their possible association with antibiotic resistance patterns of Enterococcus faecalis and Enterococcus faecium species isolated from hospital wastewater (HWW) samples of several hospitals. Methods HWW samples (200 mL) were collected from wastewater discharged from different hospitals from October 2020 to March 2021. The isolation and identification of enterococci species were performed by standard bacteriology tests and polymerase chain reaction (PCR). Antibiotic resistance was determined using the disc diffusion. The presence of various CRISPR-Cas systems was investigated by PCR. The association of the occurrence of CRISPR-Cas systems with antibiotic resistance was analyzed with appropriate statistical tests. Results In total, 85 different enterococci species were isolated and identified using phenotypic methods. The results of PCR confirmed the prevalence of 50 (58.8%) E. faecalis and 35 (41.2%) E. faecium , respectively. In total, 54 (63.5%) of 85 isolates showed the presence of CRISPR-Cas loci. The incidence of CRISPR-Cas was more common in E. faecalis . CRISPR1, CRISPR2, and CRISPR3 were present in 35 (41.2%), 47 (55.3%), and 30 (35.3%) enterococci isolates, respectively. The CRISPR-Cas positive isolates showed significant lower resistance rates against vancomycin, ampicillin, chloramphenicol, erythromycin, rifampin, teicoplanin, tetracycline, imipenem, tigecycline, and trimethoprim-sulfamethoxazole in comparison with CRISPR-Cas negative isolates. The results showed that the presence of CRISPR-Cas genes was lower in multidrug-resistant (MDR) isolates (53.1%, n = 26/49) compared to the non-MDR enterococci isolates (77.8%, n = 28/36) ( P = 0.023). Conclusion This study revealed the higher prevalence of E. faecalis than E. faecium in HWWs. Also, the lack of CRISPR-Cas genes was associated with more antibiotic resistance rates and multidrug resistance in E. faecalis and E. faecium isolates with HWW origin.
Objective and Background: SEN virus has identified as a putative new hepatitis virus coinfected with hepatitis C in thalassemic patients. The study aimed to measure viremia with SEN virus among thalassemic patients infected and non-infected with hepatitis C virus (HCV). Further, to detect the genotype D and the genetic alterations in the internal transcribed spacer domains/regions of SENV-D genotypes. Patients and Methods: A total of 200 β-thalassemic patients were included. HCV-RNA and SEN-DNA are extracting automatically. Then, amplification for SEN-DNA by conventional polymerase chain reaction (PCR) was achieved. Furthermore, HCV-RNA was amplified using real-time PCR (RT-PCR). Sequencing analysis was also done for 14 samples of amplified SENV-D DNA. Results: Out of 100 positive ELISA-anti-HCV antibodies which were definite for RT-PCR, the mean viral load in these patients was 545806 ± 1,009,799 IU/ml (1,997,176 ±3,802,206) copies/ml. There was a significant increase in SENV-D in thalassemia patients without HCV infection (59%) than those in thalassemia patients with HCV infection (44%) and healthy subjects (7%). In a sequencing study, 11 samples were infected with D genotype only while three from thalassemia patients group were coinfected with D and H genotypes. Conclusions: The occurrence rate of SEN-V DNA in thalassemia patients is very high in Iraq. Further, SEN-V-D genotype is more prevalent, and the sequences of SEN-V-D nucleotides in persons with coinfection are the same sequences of persons that have an infection with SEN-V-D alone. The most countries which have similar sequences to Iraqi SENV-D genotype sequence are Iran followed by China, Japan, Brazil, and the United States of America.
Background and objectives Peptic ulcer disease, chronic gastritis, and stomach cancer are all caused by H. pylori . The most notable drug for the treatment is the antibiotic clarithromycin, which is currently the drug of choice. H. pylori clarithromycin resistance has been associated with point mutations in 23srRNA, the most prominent of which are A2143 and A2144G . In H. pylori bacteria, methylase synthesis, macrolide-inactivating enzyme activity, and active efflux have all been found to be resistance mechanisms. The goal of the study is to determine how resistant H. pylori is to clarithromycin and what the minimum inhibitory concentration is for various antimicrobials. Furthermore, gastro-endoscopy will be performed on Iraqi patients to detect the presence of A2143G and A2144G point mutations in Helicobacter pylori infections, as diagnosed from the pyloric region and other anatomical regions. Methods One hundred fifteen samples were collected from patients strongly suspected of H. pylori infection presented for upper gastrointestinal endoscopy at Ramadi Teaching Hospitals and Private Clinics for the period from January 2020 until February 2021. Specimens were cultured on brain heart infusion agar containing various antibiotics and were incubated at 37 °C under microaerophilic conditions. For identification of H. pylori, isolates of the biochemical tests and RT-PCR assay were applied. The Epsilometer test was used in the antibiotic susceptibility testing as dependent on the CLSI standard. The Restriction Fragment Length Polymorphism technique was used to determine point mutations. Results In total, 55 (47.8%) Helicobacter pylori isolates were cultured from the 115 biopsy specimens, among which 16 (29.1%), 38 (69.1%), 20 (36.4%), and 40 (72.7%) revealed some degree of resistance to levofloxacin, clarithromycin, ciprofloxacin, and metronidazole, respectively. The frequency of A2144G and A2143 point mutations were 23 (60.5%) and 19 (50%), respectively. Conclusions According to our results , Helicobacter pylori showed high resistance to clarithromycin. Our results demonstrate the requirement for antibiotic susceptibility testing and molecular methods in selecting drug regimens.
Background and Objective:The increase in extended-spectrum β-lactamases (ESBLs) producing microbes in recent years has led to the great challenge for the clinician in the treatment. The study aims to investigate the molecular basis of ESBL encoding-resistant gene, SHV to modern β-lactams. Also, to detect gene sequencing and to compare their genetic relatedness utilizing phylogenetic analysis. Patients and Methods: A total of 100 clinical isolates are submitting after identification to phenotypic confirmatory double disk synergy technique. Polymerase chain reaction was performed for the determination of SHV gene, and sequencing analysis for the amplified gene is also achieved. Results: A total of 27 isolates, 3 (11.1%), 15 (55.55%), and 9 (33.0%) of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa were positive for a phenotypic and confirmatory test for ESBLs, respectively. SHV gene was detected only in 5 (31.25%) isolates of P. aeruginosa, 2 (12.5 %) E. coli, and 9 (56.25%) in K. pneumoniae. Conclusions: SHV gene plays an essential role in the resistance of ESBL producer isolates to new β-lactams. The sequencing of this gene revealed 98-99% compatibility range with the global standard gene in National Center of Biotechnology Information.
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