Molecular detection and PCR-RFLP analysis using Pst 1 and Alu 1 of multidrug resistant Klebsiella pneumoniae causing urinary tract infection in women in the eastern part of Bangladesh

Abstract

Klebsiella pneumoniae is the second leading causative agent of UTI. In this study, a rapid combined polymerase chain reaction and restriction fragment length polymorphism analysis was developed to identify K. pneumoniae in women, infected with urinary tract infection in the Sylhet city of Bangladesh. Analysis of 11 isolates from women at the age range of 20–55 from three different hospitals were done firstly by amplification with K. pneumoniae specific ITS primers. All of the 11 collected isolates were amplified in PCR and showed the expected 136 bp products. Then, restriction fragment length polymorphism analysis of 11 isolates were conducted after PCR amplification by 16s rRNA universal primers, followed by subsequent digestion and incubation with two restriction enzymes, Pst1 and Alu1. Seven out of 11 isolates were digested by Pst1 restriction enzymes, six isolates digested by Alu1, and while others were negative for both enzymes. Data results reveal that, women at age between 25 and 50 were digested by both enzymes. A woman aged over than 50 was negative while bellow 20 was digested by only Pst1. The results could pave the tactic for further research in the detection of K. pneumoniae from UTI infected women.

Microbiome patterns reveal the transmission of pathogenic bacteria in hilsa fish (Tenualosa ilisha) marketed for human consumption in Bangladesh

“…Some of the fish pathogens showed scattered sensitivity, whereas augmented drug resistance patterns were observed for human pathogens such as K. pneumoniae and S. aureus; in particular, one K. pneumoniae isolate showed resistance to all ten tested antibiotics. A recent study in Bangladesh found a complete resistance pattern in many K. pneumoniae isolates from the urinary tracts of infected women (Mahmudunnabi et al 2018). The drug sensitivity for other clinical bacteria, such as E. coli and P. aeruginosa, also showed a similar upwardtrending graph similar to that obtained from recently isolated clinical samples in Bangladesh .…”

“…A recent study in Bangladesh found a complete resistance pattern in many K. pneumoniae isolates from the urinary tracts of infected women (Mahmudunnabi et al . ). The drug sensitivity for other clinical bacteria, such as E. coli and P. aeruginosa , also showed a similar upward‐trending graph similar to that obtained from recently isolated clinical samples in Bangladesh (Saha et al .…”

“…E. coli ATCC 25922 and A. veronii ATCC 35624 were used as control strains. The zone of inhibition was measured using a previously described standard method (Mahmudunnabi et al 2018). Based on the CLSI guidelines (2014), the drug sensitivity patterns were divided into three distinct classes: resistant (R), intermediate resistant (IR) and sensitive (S).…”

The characterization of multidrug resistant type 1 S-fimbriated <i>Escherichia coli </i>from women with recurrent urinary tract infections (RUTIS) in Bangladesh

“…A total of 35 serial cycles of the amplification reaction were performed in a MultiGene Gradient Thermal Cycler (Labnet International Inc., USA). The PCR products were separated by electrophoresis on an agarose gel, stained with ethidium bromide solution, and visualized in a gel documentation system (8).…”

“…Drug resistance in E. coli is one of the most common barriers for treating UTI patients worldwide. However, the problem is more severe in countries like Bangladesh due to an improper tendency of frequently prescribing antibiotics for the treatment of UTIs (8). The multidrug-resistant strains of E. coli have been reported to further add to the complications in the UTI patients and decrease the effectiveness of the treatment (9,10).…”