— eT TPES sere vst expe fe myn ch Scientts Wak andthe potential impact of their werk ‘When pathogens invade human tissues, loca infection begins and biological warfare resus. Humans ate not ative observers Our resident innate immune cells (ee Bhecicn Sos Rev Vo. 25,No4 pp. 2-6) areprimed 1 telease a cascade of chemical signals in tesponse «0 infection, Local inflammation results, comtling pathogen invasion and attracting help from immune ces to axsit in microbial Gearance and tise healing the invading Pathogen has been encountered previous, an immune tmemory response can function im harmony, resulting in ox mos of our tives we exist in harmony effective pathogen eradication, More severe infections may SAMA E Fert microbes Soon after bith ou sterile Wwigera ever, which poets against the pathogen. These Aigestive tract becomes colonised with host mmune responses are carefully regulated and aimed at ject microorganisms. The complex microbial infection survival and rewrn to normal function innacaee ecosystem that develops inthe gut has major While mostimmune espns ate effective at combating wa benefits forhuman health pomotingeffcient the infection, some severe infections can elicit an immune Dagens Ligestion offood and developmentofaheslthy response that causes ancontolled inflammation. This immune sytem, Hover, some microbes — response can spread widely throughout the body in classed as pathogens — can pose a significant threat w humans because the bloodstream, rapidly becoming life-threatening as they are able to cause disease, Some pathogens have develope the abilty each organ in the boxy becomes inflamed ans ceases 10 to ofercome human defences. When they gain entry to the body they function. Known s sepsis, this complication ofthe human proliferate and secrete chemicals that are tori to man. Other pathogens immune response to infection isa major global heatheare may be normally harmless microbes present on body surfaces such asthe challenge, responsible for mllions of deaths per year I is skin or inthe gut. They may enter the body during disease, for example not clear why some people develop sepsis while others do elective immanity, oF following inary such as a surface wound, Hence, ot, but the spe of microorganism, previous exposure to 4rdelicate balance exins between man and microbes that is crucial in antibiotics, presence of otherlinescs and genetics al play _llUOe ee Samples rom postive Sonplesfom postive bleed Samples from pov blood culture bots pated Dioodcuiture bates ae elturebotes plated onto sold onto sola auture media coming a vere removed fom the incubetor_“eulturemeciatofelp further of dacs impregnated wth common antimicrobial. andaiodedundera microscope ““ilenfation fesione droge tohelp deewrnne te ike sens, "er early cauiicaton presnt Common tater andveritarcepateme of he growing mire. and entation further -2aaysto complete Comment takes further few das of eutore Figure + Procedure for clturing blood samples to identity the bacterium or fungus causing sepsie November 2013presenti the patient's blodstream willbe induced ino the blood cukure bots, making subsequent pathogen caulure mote pioblemav In an atiempe © prevent this Problem, fis tecommendd that al lood culture samples be talon hefore giving antibiotics, making blood cultures 8 ‘ruil part ofthe emergency cate of patient with sepsis The clinical value of blood cultures in tweaking tepis {is when 4 pathogea is dentfied. Microbial species can be (0) Neterals sod the POR machine Peet we COG Baan Terpi OWA Pane (2) Thoma sages ote ptmerese cain reacton Hest 995°0 Da stance | cea 0-5 2S. nesting ‘nd eoorg balou ert men copes Sime ongna ONA Primers te tangle sane ‘enplonerany Sierra Im Heat io720 in each yc the DNA pola eure otonce number oops of tamplemorary ONA sands, BeDNAtompate ls ring tom fe primes oes “wo cops of ONA sans Figure The polymerase chain reaction 18 “Mentfied by observing characteristics uch as sizeand shape Lunde the microscope. Knowledge of microbial speces can help the weatng clinician stat wo reine the anionic used “The identifiation ofthe pathogen speces can help race ‘he patient's exposure to potent bread spectrum antibion, Considering the time taken for culture dagoosis, clinical scientists have been draen to recent advances in technology ‘wallow erly diagnosis and pathogen Mlentification Solutions from amplifying the problem actria coazain DNA which, asin humans, encodes crcl informatio relating to form and function. Unlike human DNA, the DNA ofacteria snot contained within a nuclear eswelope (ee pp 20-21). Another difference isthat bacterial DNA contains a unique segment of sequences chat encode genetic information tlating to their ibosomnal RNA. This Segment is ony furl in bacterial DNA ad is remarkably tonserved from one generation 1o the next. Sight sequence ‘variations inthis segment are specific to diferent bacterial species. racial these sequences are not fund in mammalian DNA and so could provide key amgets for devecting and ‘emying bacterial species present inhuman samples However, in cases of sepsis, pathogen detection will be Aifeuk Human blood contains lage amount of human DNA, mainly in cicustingimmune cells. Pathogen numbers are very lve inthe blood of patents with sepsis. Finding microbial DNA in ths setting would be lke hunting for 8 slog gas blade in a lange playing el Technical advances provide an elegant solution to this problem. The polymerase chain reaction (PCR) is technique that will amplify a specific segment of DNA, snabling a particular vequence o be ented even in the presence of high concentrations of other DNA (ee Figure 2} Amplification ofa unique bacterial DNA segment means that even low quantities of bacterial DNA can be Mentied fn the presence of human DNA, This allows pathogen to be rapidly Mentified fom 3 patient’ blood samples PCR technology provies several portunities in sepsis diagnosis. simple diagnostic west could be developed to ‘determine rapidly whetherbactrial or fungal DNA ipreseat ina blood sample. Howeves, itis curently not possible to deliver comprehensive rapid agnostic vest hat contains Information about a detected pathogens likely anobiotic resistance, Although carried by pathogen DNA, most Antimicrobial resistance information is exttemely complex tnd the mechanism of resistance isnot full understood. ‘Despite thissignificant limitation, commercial sborstory CR pathogen diagnostic tests ate now emerging aimed aC patients with sepsis, A number have gained Luropean regulatory approval but none has heen tested to date In Carefully designed clinical tal to determine whether they ae effective at improving the cae of patente with sepa, DNA sequencing: a diagnostic future DNAencodes the genetcinstuctionsusedin thedevelopment and function of al living organisms, iclaing baer, Fungi and some viruses, Genet information fence a6 bloga eres eensequences of nucleotides. The technical proceses of DNA sequencing have been developed to uncover the precise ‘order ofthe ruleaties in DNA, Complex mathematics association with powerful computers canbe used to break tenetic codes by iemifying particular geneticsequences and Ssicitingthem with the development of form and function ofan organism, Ia the case of DNA from microorganisms, sequencing can be used to identity the microbe present. its species a8 ‘well as is likey ability to cause disease adi probable tesistance 10 different antimicrobial drugs, Modern Sequencing technologies are becoming more powerful in terms of their precision and speed, with an associated HODDER averse 4) decline inthe size of instrumentation and cost. 1s likely lover the next decade that capi laboratory diagnostics in ‘microbiology willbe realised using the next generation of DNA sequencing technologies, which hold real potential for Arvin significant improvement inpatient careforcommon lie cheatening conditions such a sepa Intheinterim,itisimportantt testclnically and develop farther the mote limied current commercial approaches to pathogen detection and Henifcation chat ae gaining "uropean regulatory approval. These echniques will prose Important Knowledge about haw diagnostic information delivered rapidly to the clinician i Ikely to impact on tweatinen decisions and outcome in sep. In addin, these liical studies can be used to inform the reguirements and configuration of net generation sequencing technologies, For instance, what information about a microbe will be ‘most useful in delivering beter are fo patients with sepsis! Aid, elated to thie, should these rapid diagnostic tests be developed on a microchip atthe pointof ate, or should they be locate ina large centralised diagnostic laboratory? Getting the most from your magazine? 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