5f30e772-5406-4c40-aeb7-84f62ccfcbe0
In Iran, Klebsiella pneumoniae is one of the dangerous bacteria in nosocomial infections. In Klebsiella pneumonia, antibiotic resistance is very high and with the production of ESBL, it is resistant to almost all common antibiotics. In our project, we have tried to isolate and replicate specific phages against this bacterium so that we can use them in next phage therapy projects. Genomes are uploading for using by other researchers and improving phage science.
In Iran, Klebsiella pneumoniae is one of the most dangerous bacteria in nosocomial infections. In Klebsiella pneumonia, antibiotic resistance is very high and with the production of ESBL, it is resistant to almost all common antibiotics. In our project, we have tried to isolate and replicate specific phages against this bacterium so that we can use them in the next phage therapy projects. In this project, sampling was performed of the wound from a 62-year-old patient who had a severe bedsore. Then, after passing the sewage of Bu-Ali Hospital in Qazvin through a 0.22 µm filter, was added Klebsiella pneumoniae, and the desired phage was able to destroy Klebsiella pneumoniae. After using the plaque assay technique and purification, we tacked electron microscopic photos. The phage is belonging to siphoviridae family with dimensions (Capsid=60 nm, Tail=130nm). Phage DNA was extracted using Phage DNA Isolation Kit (NORGEN). Genomic DNA libraries are prepared using the Nextera XT Library Prep Kit (Illumina, San Diego, USA) following the manufacturer's protocol with the following modifications: input DNA is increased 2-fold, and PCR elongation time is increased to 45 s. DNA quantification and library preparation are carried out on a Hamilton Microlab STAR automated liquid handling system (Hamilton Bonaduz AG, Switzerland). Pooled libraries are quantified using the Kapa Biosystems Library Quantification Kit for Illumina. Libraries are sequenced using Illumina sequencer (HiSeq/NovaSeq) using a 250bp paired-end protocol. Reads were adapter trimmed using Trimmomatic 0.30 with a sliding window quality cutoff of Q15. De novo assembly was performed on samples using SPAdes version 3.7, and contigs were annotated using Prokka 1.11. Genomes are uploaded for use by other researchers and improve phage science.
In Iran, Klebsiella pneumoniae is one of the most dangerous bacteria in nosocomial infections. In Klebsiella pneumonia, antibiotic resistance is very high and with the production of ESBL, it is resistant to almost all common antibiotics. In our project, we have tried to isolate and replicate specific phages against this bacterium so that we can use them in the next phage therapy projects. In this project, sampling was performed of the wound from a 62-year-old patient who had a severe bedsore. Then, after passing the sewage of Bu-Ali Hospital in Qazvin through a 0.22 µm filter, was added Klebsiella pneumoniae, and the desired phage was able to destroy Klebsiella pneumoniae. After using the plaque assay technique and purification, we tacked electron microscopic photos. The phage is belonging to siphoviridae family with dimensions (Capsid=60 nm, Tail=130nm). Phage DNA was extracted using Phage DNA Isolation Kit (NORGEN). Genomic DNA libraries are prepared using the Nextera XT Library Prep Kit (Illumina, San Diego, USA) following the manufacturer's protocol with the following modifications: input DNA is increased 2-fold, and PCR elongation time is increased to 45 s. DNA quantification and library preparation are carried out on a Hamilton Microlab STAR automated liquid handling system (Hamilton Bonaduz AG, Switzerland). Pooled libraries are quantified using the Kapa Biosystems Library Quantification Kit for Illumina. Libraries are sequenced using Illumina sequencer (HiSeq/NovaSeq) using a 250bp paired-end protocol. Reads were adapter trimmed using Trimmomatic 0.30 with a sliding window quality cutoff of Q15. De novo assembly was performed on samples using SPAdes version 3.7, and contigs were annotated using Prokka 1.11. Genomes are uploaded for use by other researchers and improve phage science.