Genetic fingerprinting and profile analysis of virulence genes in XDR clinical isolates of Klebsiella Pneumoniae.

OBJECTIVE: Klebsiella pneumoniae, a leading cause of hospital- and community-acquired infections, requires the acquisition of drug resistance and virulence for survival in evading the host's immune responses. The investigation looks at the molecular diversity of 30 clinical isolates using rep-PCR DNA fingerprinting and the association between drug resistance and virulence genes. MATERIALS AND METHODS: Isolates were cultured using basic bacteriological techniques, while antimicrobial susceptibility test and ID confirmation were made by Vitek 2 Compact Automated System (BioMerieux, Marcy L'Etoile, France). DNA used for PCR reactions was extracted with Qiagen DNA extraction kit according to the manufacturer's guidelines, while molecular genotyping by REP-PCR was carried out according to previously described methods. Some virulence genes and capsular serotypes, ybtS, mrkD, entB, rmpA, K2, Kfu, alls, iutA, and magA, were amplified by multiplex PCR. RESULTS: The majority (60%) of the isolates were MDR, others were XDR (37%) and susceptible strains (3%). Additionally, resistance was high (73%) for meropenem and lower (40%) for imipenem, while 97% were susceptible to ertapenem and azithromycin. DNA fingerprinting by rep-PCR showed polymorphic loci sizes that ranged from 100-2000 bp-, and phylogenetic analysis placed the isolates in seven clades with 40-96%. There was a 75% phylogenetic relatedness to antimicrobial resistance, but no specific pattern of relatedness was seen between virulence genes for the K. pneumoniae strain. Regulator mucoid phenotype A (rmpA) gene was not amplified in any of the isolates. CONCLUSIONS: This study further confirms the presence of both MDR and XDR K. pneumonia strains. Isolates exhibited genetic diversity in phylogenetic patterns and carriage of virulence with relatedness by susceptibility to antimicrobials. Therefore, circulation could pose a risk to public health.

The malaria burden: A look at 3 years outpatient malaria clinic visits in a university community town in Southeast of Nigeria.

BACKGROUND: One of the Nigerian vision 2020 National Malaria strategic plans is control and subsequent eradication of malaria. The present report looks at outpatient malaria clinic visits for a 3-year period with a view of ascertaining whether control measures put in place over decades are being reflected in the decline of the disease. METHODS: A retrospective study was conducted at Nsukka in the southeast Nigeria using a cluster sampling method for the selection of health care facilities. Collected data included patient demography, number attendees, and the levels of parasitemia. The "Plus System Scale" was used for the grouping of detected levels of Plasmodium parasites in the blood samples and data were analyzed using SPSS (version 23). RESULTS: A total of 9,531 outpatient malaria clinic visits which consisted of females (67.5%) and males (32.5%) were used for the report. The difference in the number of males and female malaria clinic attendees was statistically significant [P < 0.05]. Examined blood samples showed 87.25% were positive with Plasmodium falciparum parasites with various levels of parasitemia. There were also negative Plasmodium parasites blood samples with mean scores of 67 (±22.62), 92.63 (±9.97), and 353 (±179.6) for years 2013, 2014, and 2015, respectively. Most (43.47%) of the patients were in the age group of 21-30 and while parasitemia was seen to be higher in this group (21-30). CONCLUSION: The incidence of malaria in the region of this study is still high despite the effects made at reducing the scourge of the disease and would need timely intervention.