Evaluation of single and double-locus real-time PCR assays for methicillin-resistant Staphylococcus aureus (MRSA) surveillance

<p>Abstract</p> <p>Background</p> <p>Methicillin-resistant <it>Staphylococcus aureus </it>(MRSA) is a human pathogen, representing an infection control challenge. Conventional MRSA screening takes up to three days, therefore development of rapid detection is essential. Real time-PCR (rt-PCR) is the fastest method fulfilling this task. All currently published or commercially available rt-PCR MRSA assays relay on single or double-locus detection. Double-locus assays are based on simultaneous detection of <it>mecA </it>gene and a <it>S. aureus</it>-specific gene. Such assays cannot be applied on clinical samples, which often contain both coagulase-negative staphylococci (CoNS) and <it>S. aureus</it>, either of which can carry <it>mecA</it>. Single-locus assays are based on detection of the staphylococcal cassette chromosome <it>mec </it>(SCC<it>mec</it>) element and the <it>S. aureus</it>-specific <it>orfX </it>gene, assuming that it is equivalent to <it>mecA </it>detection.</p> <p>Findings</p> <p>Parallel evaluation of several published single and double-locus rt-PCR MRSA assays of 150 pure culture strains, followed by analysis of 460 swab-derived clinical samples which included standard identification, susceptibility testing, followed by PCR detection of staphylococcal suspected isolates and in-PCR mixed bacterial populations analysis indicated the following findings.</p> <p>Pure cultures analysis indicated that one of the single-locus assay had very high prevalence of false positives (Positive predictive value = 77.8%) and was excluded from further analysis. Analysis of 460 swab-derived samples indicated that the second single-locus assay misidentified 16 out of 219 MRSA's and 13 out of 90 methicillin-sensitive <it>S</it>. <it>aureus</it>'s (MSSA) were misidentified as MRSA's. The double-locus detection assay misidentified 55 out of 90 MSSA's. 46 MSSA containing samples were misidentified as MRSA and 9 as other than <it>S. aureus </it>ending with low positive predicted value (<85%) and very low specificity (<62%).</p> <p>Conclusion</p> <p>The results indicate that high prevalence of false-positive and false-negative reactions occurs in such assays.</p

The influence of a family\u27s lifestyle on the physical activity and healthy diet of preschool children

Diplomsko delo z naslovom Pomen družinskega sloga za gibalno športno aktivnost in zdravo prehrano predšolskih otrok je sestavljeno iz dveh delov, iz teoretičnega in empiričnega dela. V teoretičnem delu je najprej nekaj besed namenjenih temu, da je zelo pomembno razmerje med vnosom in porabo energije in kako se je življenjski slog družin bistveno spremenil, odkar je med nami tehnologija. Nato smo predstavili življenjski slog in kako stres v življenju v veliki meri vpliva na življenjski slog posameznika. V naslednjem poglavju smo opredelili družino in kaj vse spada k družinskemu okolju. Opisali smo tudi pomen otrok v družini in kako so otroci povsem odvisni od staršev. Opredelili smo tudi gibalno športno aktivnost nasploh in v družini, vključevanje otrok v le-to ter kaj so posledice premajhne športne aktivnosti. V empiričnem delu smo s pomočjo table prikazali odgovore, ki smo jih pridobili na podlagi anketnega vprašalnika. Le-ta je zajemal vprašanja, ki so se nanašali na pogostost, pomembnost in rednost zdrave prehrane in športnih gibalnih aktivnosti v družinah. Vse anketirane osebe so bili starši predšolskih otrok. Vsako tabelo smo interpretirali na podlagi pridobljenih rezultatov. Prišli smo do ugotovitve, da ima življenjski slog na prehrano in gibalno športno aktivnost predšolskih otrok velik vpliv, saj so otroci v veliki meri odvisni od prehrane in gibalno športnih aktivnosti, ki jim jih nudijo starši.The diploma thesis with the title The Importance of a Family\u27s Lifestyle for the Physical Activity and Healthy Diet of Preschool Children consists of two parts, the first one being theoretical and the second one empirical. In the theoretical part, a few words are first devoted to the fact that the relationship between energy intake and consumption is very important and how the lifestyle of families has changed significantly since technology has entered our lives. We then described what lifestyle is and how stress greatly affects an individual’s lifestyle. In the next chapter, we defined the term family and discussed what a family environment encompasses. We also described the importance of children in a family and how children are completely dependent on their parents. We also defined physical activity in general and in the family, the inclusion of children in families and what the consequences of insufficient sports activity are. In the empirical part, we used a table to show the answers we obtained on the basis of our questionnaire. The questionnaire included questions related to the frequency, importance and regularity of healthy eating and physical activity in families. All respondents were parents of preschool children. Each table was interpreted on the basis of the obtained results. We concluded that the lifestyle of nutrition and physical activity of preschool children is of great importance, as children are largely dependent on the diet and physical activities offered to them by their parents

A global point prevalence survey of antimicrobial use in neonatal intensive care units : The no-more-antibiotics and resistance (NO-MAS-R) study

Background: Global assessment of antimicrobial agents prescribed to infants in the neonatal intensive care unit (NICU) may inform antimicrobial stewardship efforts. Methods: We conducted a one-day global point prevalence study of all antimicrobials provided to NICU infants. Demographic, clinical, and microbiologic data were obtained including NICU level, census, birth weight, gestational/chronologic age, diagnoses, antimicrobial therapy (reason for use; length of therapy), antimicrobial stewardship program (ASP), and 30-day in-hospital mortality. Findings: On July 1, 2019, 26% of infants (580/2,265; range, 0-100%; median gestational age, 33 weeks; median birth weight, 1800 g) in 84 NICUs (51, high-income; 33, low-to-middle income) from 29 countries (14, high-income; 15, low-to-middle income) in five continents received &gt;= 1 antimicrobial agent (92%, antibacterial; 19%, antifungal; 4%, antiviral). The most common reasons for antibiotic therapy were "rule-out" sepsis (32%) and "culture-negative" sepsis (16%) with ampicillin (40%), gentamicin (35%), amikacin (19%), vancomycin (15%), and meropenem (9%) used most frequently. For definitive treatment of presumed/confirmed infection, vancomycin (26%), amikacin (20%), and meropenem (16%) were the most prescribed agents. Length of therapy for culture-positive and "culture-negative" infections was 12 days (median; IQR, 8-14) and 7 days (median; IQR, 5-10), respectively. Mortality was 6% (42%, infection-related). An NICU ASP was associated with lower rate of antibiotic utilization (p = 0.02). Interpretation: Global NICU antibiotic use was frequent and prolonged regardless of culture results. NICU-specific ASPs were associated with lower antibiotic utilization rates, suggesting the need for their implementation worldwide