Construction, Cloning, and Expression of CagA Recombinant Protein of Helicobacter pylori.

BACKGROUND: This study aimed to assess construction and expression of CagA recombinant protein of Helicobacter pylori (H. pylori) in Escherichia coli (E. coli) BL21. METHODS: Bioinformatics was used in designing the desired gene by Gene Runner. Next, the construct was subcloned to pET21b vector and this process was confirmed by Polymerase Chain Reaction (PCR), enzyme digestion and sequencing techniques. Then, it was cloned in the Escherichia coli BL21 as an expression host. Expression of protein was verified using sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting technique. For purification of the protein, the Ni-NTA column was used. Protein concentration was determined by the Bicinchoninic Acid Protein Assay Kit (Parstoos). Finally, Western blotting was performed using CagA antibodies and normal human serum for determining immunogenicity feature with human antiserum. RESULTS: According to the results of the present study, CagA construct was cloned into the pET21b vector and after confirmation and cloning in host expression, recombinant protein with the size of 38 kDa was successfully expressed and purified. The recombinant CagA protein showed immunogenicity characteristics with human antiserum. CONCLUSION: In conclusion, only 5'-end of recombinant protein CagA with high immunogenicity effects was successfully constructed, cloned and expressed. Also, CagA recombinant protein showed good immunogenicity activity with human antiserum.

Aspergillus coinfection among patients with pulmonary tuberculosis in Asia and Africa countries; A systematic review and meta-analysis of cross-sectional studies.

Progress of the disease and prolonged treatment with antibiotics or immunosuppressive agents makes tuberculosis patients susceptible to fungal infections. This study aimed to determine the prevalence of pulmonary Aspergillus coinfection among patients with pulmonary tuberculosis in Asia and Africa. The present review of cross-sectional studies was conducted on the prevalence of pulmonary Aspergillus coinfection among patients with pulmonary tuberculosis according to the PRISMA Protocol. Literatures published online in English from January 2001 to March 2019 via key databases such as Web of Science, MEDLINE, PubMed, Scopus, and Cochrane Library were searched. The used MeSH and non-MeSH keywords were; "pulmonary fungal", "pulmonary coinfection", OR "Pulmonary mycosis", "pulmonary fungal infections/agents", OR "Polymicrobial infection", OR "Secondary infection", OR "Mixed infections", "pulmonary aspergillosis", "fungi coinfection", "Fungal co-colonization", AND "pulmonary tuberculosis", OR "pulmonary TB", AND "Asia" AND "Africa". Finally, data analyzed using Comprehensive Meta-Analysis software (CMA). The combined Aspergillus coinfection among patients with pulmonary tuberculosis was 15.4% (95% CI: 11.4-20.5), Q = 105.8 and Z = 9.57 in Asia and Africa. The most frequency of Aspergillus spp. was related to A. fumigatus with a combined prevalence of 57.6%. Most of the studies included in the present review showed a higher Aspergillus coinfection in the age group of 40 years and higher. Also, the existence of a correlation between increasing age and Aspergillus coinfection was reported (p < 0.05). The present review showed a high combined Aspergillus coinfection among patients with pulmonary tuberculosis in Asia and Africa. Also, amongst the Aspergillus spp., the most frequent was related to A. fumigatus.

Prevalence of Integrons and Antibiotic Resistance Pattern in Acinetobacter baumannii Isolated from Clinical Samples of Iranian Patients: A Systematic Review and Meta-analysis.

BACKGROUND: Acinetobacter baumannii is an important opportunistic nosocomial pathogen. Class 1 integrons in A. baumannii plays a significant role in antibiotic resistance. Therefore, this study aimed to investigate the prevalence of integrons and antibiotic resistance pattern in A. baumannii isolated from clinical samples of Iranian patients. METHODS: The Medical Subject Headings (MeSH) and the keywords with the help of Boolean operators ("AND" or "OR") were used alone or in combination to conduct the search. The searching process was conducted in the Web of Science, PubMed, Cochrane Library, Scopus, and Google Scholar databases and, also Iranian databases. The search was restricted to relevant English and Persian cross-sectional publications reporting the prevalence of Int1 in A. baumannii isolated from clinical samples from 1 January 2000 to 31 December 2018. The data were analyzed using Comprehensive Meta-Analysis software. Regarding the heterogeneity of studies, the random effects model was used. Cochrane Q and I2 tests was used to evaluate statistical heterogeneity between the studies. RESULTS: Fifteen studies were included in the analysis. The combined prevalence of class 1 integrons in A. baumannii was 55.2% (95% CI: 44.8-65.1). The pooled prevalence of MDR A. baumannii isolates was 68.1%. The highest resistance belonged to Aztreonam, followed by Ciprofloxacin, and Ceftazidime with a resistance rate of 97.6%, 92.8%, and 91.6%, respectively. Tobramycin was reported as an effective antibiotic. CONCLUSIONS: The present study reported an alarmingly high prevalence of class 1 Integrons, and MDR isolates of A. baumannii recovered from clinical samples that should be considered.