Ob koncu lanskega leta je Zdravstveni svet Republike Slovenije potrdil program preprečevanja zgodnjih invazivnih okužb novorojenčkov, povzročenih s streptokokom skupine B, ki vključuje univerzalno ...presejanje nosečnic v 35.–37. tednu nosečnosti. V prispevku smo pregledali različne diagnostične metode presejanja in dejavnike, ki pomembno vplivajo na njihovo uspešnost tako v ginekološki ambulanti kakor tudi v mikrobiološkem laboratoriju. Bralca opozarjamo na pravilnost odvzema kužnine, mu predstavimo izbrano strategijo testiranja s kombinacijo obogatene kulture in molekularnega testiranja ter mu ponujamo seznam registriranih molekularnih testov, primernih za testiranje. V zadnjem delu prispevka razpravljamo o pomenu hipervirulentnega klona CC-17, ki povzroča večino invazivnih okužb novorojenčkov v Sloveniji, in o metodah, s katerimi ga prepoznamo.
The purpose of our study was to investigate methods of short-term storage that allow preservation, transport and retrieval of genetic information contained in honeybee queen's spermatheca. Genotyping ...of the honeybee colony requires well ahead planned sample collection, depending on the type of data to be acquired. Sampling and genotyping of spermatheca's content instead of individual offspring is timesaving, allowing answers to the questions related to patriline composition immediately after mating. Such procedure is also cheaper and less error prone. For preservation either Allprotect Tissue Reagent (Qiagen) or absolute ethanol were used. Conditions during transportation were simulated by keeping samples 6-8 days at room temperature. Six different storing conditions of spermathecas were tested, complemented with two DNA extraction methods. We have analysed the concentration of DNA, RNA, and proteins in DNA extracts. We also analysed how strongly the DNA is subjected to fragmentation (through amplification of genetic markers ANT2 and tRNA
-COX2) and whether the quality of the extracted DNA is suitable for microsatellite (MS) analysis. Then, we tested the usage of spermatheca as a source of patriline composition in an experiment with three instrumentally inseminated virgin queens and performed MS analysis of the extracted DNA from each spermatheca, as well as queens' and drones' tissue. Our results show that median DNA concentration from spermathecas excised prior the storage, regardless of the storing condition and DNA extraction method, were generally lower than median DNA concentration obtained from spermathecas dissected from the whole queens after the storage. Despite the differences in DNA yield from the samples subjected to different storing conditions there was no significant effect of storage method or the DNA extraction method on the amplification success, although fewer samples stored in EtOH amplified successfully in comparison to ATR storing reagent. However, we recommend EtOH as a storing reagent due to its availability, low price, simplicity in usage in the field and in the laboratory, and capability of good preservation of the samples for DNA analysis during transport at room temperature.
