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    Evaluation and validation of room temperature biospecimens transportation and storage technologies as an alternative cost effective solution to cold chain logistics and storage within biobanking and/or diagnostics

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    Thesis (MSc)--Stellenbosch University, 2017ENGLISH ABSTRACT : Background: Cold chain management (CCM) is an important aspect of biobanking operation. However challenges such as constant power failure, limited access to dry ice transport and storage of human samples collected at various sites all over the world or at difficult out of reach places. and liquid nitrogen, transport logistics and courier delays especially in Africa becomes a major challenge. Ensuring samples are maintained at the proper temperature throughout all processes is imperative to maximal long term viability and usability. Thus we consider room temperature storage (RTS) technologies as an innovative, cost effective and green alternative to cold chain logistics. Methods: Various room temperature storage technologies were evaluated for the stabilization and storage of whole blood DNA and RNA, buffy coat, genomic DNA and urine DNA. The stabilizers include the Biomatrica liquid gard technology and dry matrix technology as well as DNAgenotek Hemagene buffy-coat stabilizers, Paxgene RNA and Norgen urine tubes. Samples were stored with and without a stabilizer under different temperature conditions namely room temperature, 45oC,-80oC, -20oC and liquid nitrogen (- 196oC) over different time periods to determine effect on sample integrity and quality. At the end of each time point DNA/RNA was extracted and the integrity of the samples determined by assessing the concentration, purity and integrity. Further downstream analysis such as polymerase chain reaction (PCR), quantitative real time PCR and DNA sequencing was conducted. In addition, a shipping cost analysis between satellite sites in African and our biobank was done to compare frozen and room temperature shipping. Results The study results show that sample integrity/quality for biospecimens stored at room temperature with stabilizers were comparable and more cost effective than cold chain storage systems. In addition some stabilizers showed better stabilizing properties than others. Conclusion: Room temperature storage provides an innovative and cost effective method of storage and shipping to cold chain management systems (CCM). Green technologies forms a small part of biobanking operations however its results would be beneficial as low energy options for biobanking are particular critical in low resource settings which have infrastructural challenges. In turn, it would also be a more cost-effective option for the transport and storage of human samples collected at various sites all over the world or at difficult out of reach places.AFRIKAANSE OPSOMMING : Agtergrond: Koue ketting bestuur (KKB) is 'n belangrike aspek van Biobank bedrywighede, maar uitdagings soos konstante kragonderbrekings, beperkte toegang tot droë ys en vloeibare stikstof, vervoer logistiek en koerier vertragings veral in Afrika is 'n groot uitdaging. Om te verseker dat monsters in stand gehou word by die regte temperatuur, in die hele proses, is dit noodsaaklik om lang lewensvatbaarheid en bruikbaarheid van monsters te maksimiseer. Dus kyk ons na kamertemperatuur stoor tegnologie as 'n innoverende, koste-effektiewe en groen alternatief vir koue ketting logistiek. Metodes: Verskeie kamertemperatuur stoor tegnologie is geëvalueer vir die stabilisering en stoor van heelbloed DNA en RNA, “buffy coat”, genomiese DNA en uriene DNA. Die stabiliseerders sluit in die Biomatrica vloeibare beskerm-tegnologie en droë matriks tegnologie asook DNAgenotek, Hemagene “buffy coat” stabiliseerders, Paxgene RNA en Norgen urienbuise. Monsters is gestoor met en sonder 'n stabiliseerder by verskillende temperature naamlik kamertemperatuur, 45oC, -80oC, -20oC en in vloeibare stikstof (- 196oC) oor verskillende tydperke om die uitwerking op monster integriteit en kwaliteit te bepaal. Aan die einde van elke tydperk is DNA / RNA geisoleer en die integriteit, kwaliteit en konsentrasie van elke monsters is bepaal en geëvalueer. Verdere stroomaf ontleding soos Polimerase Kettingreaksie (PKR), kwantitatiewe “real-time” PKR en DNA volgordebepaling is gedoen. Hierby is 'n koste-ontleding tussen satelliet gebiede in Afrika en ons Biobank gedoen om bevrore en kamertemperatuur monsters wat aangestuur is na die lande te vergelyk. Resultate: Die studie wys dat die integriteit/kwaliteit van monsters wat by kamer temperatuur gestoor was in stabiliseerders, vergelykbaar en meer koste-doeltreffend as koueketting stoor stelsels was. Daarbenewens het 'n paar stabiliseerders beter stabiliserende eienskappe as ander getoon. Gevolgtrekking: Kamertemperatuur berging bied 'n innoverende en meer kosteeffektiewe metode vir die stoor en stuur van monsters as koue ketting bestuurstelsels. Groen tegnologie vorm 'n kleindeel van Biobank bedrywighede, maar die resultate sal egter voordelig wees as lae energie-opsies vir Biobank bedrywighede, en is besonder krities in lae hulpbron instellings wat uitdagings met infrastruktuur ervaar. Op sy beurt, sou dit ook 'n meer koste-effektiewe opsie wees vir die vervoer en berging van menslike monsters na verskillende plekke oor die wêreld of in moeilike bereikbare plekke
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