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Oundo, V.

Publications and source records attributed to Oundo, V..

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A 7-member SNP Assay on the iPlex MassARRAY Platform Provides a Rapid and Affordable Alternative to Typing Major African Staphylococcus aureus Types

BackgroundData on the clonal distribution of Staphylococcus aureus in Africa is scanty, partly due to high costs and long turnaround times imposed by conventional genotyping methods such as spa and multilocus sequence typing (MLST) warranting the need for alternative typing approaches. This study applied and evaluated the accuracy, cost and time of using iPlex massARRAY genotyping method on Kenyan staphylococcal isolates.\n\nMethodsFifty four clinical S. aureus isolates from three counties were characterized using iPlex massARRAY, spa and MLST typing methods. Ten Single Nucleotide Polymorphisms (SNPs) from the S. aureus MLST database were assessed by iPlex massARRAY.\n\nResultsThe iPlex massARRAY assay grouped the isolates into 14 SNP genotypes with 9/10 SNPs interrogated showing high detection rates (average 89%). spa and MLST typing revealed 22 spa types and 21 STs that displayed unique regional distribution. spa type t355 (ST152) was the dominant type and t2029 and t037 (ST 241) were observed among MRSA strains. MassARRAY showed 83% and 82% accuracy against spa and MLST typing respectively in isolate classification. Moreover, massARRAY identified all MRSA strains and a novel spa type. MassARRAY had reduced turnaround time (<12 hrs) compared to spa (3 days) and MLST (20 days) typing. The iPlex massARRAY cost approximately 18 USD compared to spa (30 USD) and MLST (126 USD) typing based on consumable costs/isolate.\n\nConclusionUpon validation with a larger collection of isolates, iPlex massARRAY could provide a faster, more affordable and fairly accurate method of resolving African S.aureus isolates especially in large surveillance studies.

microbiology

Strain typing and characterization of virulence genes in clinical Staphylococcus aureus isolates from Kenya

Staphylococcusaureus strain typing is an important surveillance tool as particular strains have been associated with virulence and community and hospital acquired MRSA outbreaks globally. This study sought to determine the circulating strain types of S.aureus in Kenya and establish the virulence genes among the strains. Clinical S.aureus isolates from 3 hospitals in Kenya were sequenced on the Illumina Miseq and genomes assembled and annotated on PATRIC. Results demonstrated great diversity among the isolates with identification of 6 distinct CC (8,22,15,80,121,152), 8 ST types (8, 15, 22,80,121,152,241, 1633) and 8 spa types (t005, t037, t064, t084, t233, t2029, t272,t355). Novel STs (4705, 4707) and a novel spa type (t17826) were identified. The most prominent clonal complex was CC 152 comprised of only MSSA. A majority of MRSA isolates (3/4) typed to ST 241, CC8. One MRSA isolate typed to a novel ST 4705. All isolates were screened for a panel of 56 known virulence genes (19 adhesins, 9 hemolysins, 5 immune evasion proteins, 6 exo-enzymes and 19 toxins). 9 toxin genes were detected among the isolates with CC8 isolates having the highest numbers of toxin genes. An MSSA isolate (CC8) from a severe burn infection had the highest number of toxin genes (5). All MRSA isolates (CC8) had only 2 toxins, SEK and SEQ, whereas a majority of the MSSA isolates either had 0 or [&ge;]2 toxins. SEK+SEQ and TSST-1+SEB+SEL toxin combinations were observed among patients whose disease resulted in hospitalization, an indicator of severe infections. This study confirms the highly heterogeneous S.aureus population in Kenya. MSSA appear to have the potential of accumulating more toxin genes than MRSA. This co-occurrence of major toxin genes, some associated with MRSA, highlights the potential risks of outbreaks of highly virulent MRSA infections which would pose treatment challenges.

microbiology