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  <controlfield tag="008">190826s2017    th uu m rtt   0|  a1eng d</controlfield>
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    <subfield code="a">AIT Thesis no.FB-17-02</subfield>
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    <subfield code="a">Jayanath, Nileththi Yasendra</subfield>
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    <subfield code="a">Development of an electrochemical device capable of loop mediated isothermal amplification (LAMP) to detect hepatitis B</subfield>
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    <subfield code="a">Pathum Thani, Thailand :</subfield>
    <subfield code="b">Asian Institute of Technology,</subfield>
    <subfield code="c">2017</subfield>
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    <subfield code="a">Thesis ;</subfield>
    <subfield code="v">no.FB-17-02</subfield>
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    <subfield code="a">A thesis submitted in partial fulfillment of the requirements for the  degree of Master of Science in Food Engineering and Bioprocess Technology,  School of Environment, Resources and Development</subfield>
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    <subfield code="a">Thesis (M. Sc.) - Asian Institute of Technology, 2017</subfield>
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    <subfield code="a">A sensitive, simple and inexpensive technique for point - of - care (POC) diagnosis of hepatitis  B (HBV) is  highly  desirable majorly  due  to  the  lack  of  advanced  medical  and  diagnostic facilities in some endemic areas . The objective of this study was to  develop a low - cost and  simple  device  based  on  Loop - mediated  isothermal  amplification  (LAMP)  of  DNA  and  electrochemical  measurement  for  rapid  detection  of  HBV. The  electrochemical  mediato r  methylene  blue  (MB)  was  intercalated  into  the  LAMP  amplicons  and  square  wave  voltammetry (SWV) signals were processed by a hand - held potentiostat. SWV response of  MB  was  studied  with  different  concentrations.  LAMP  was  optimized  for  temperature  and  MB con centrations for real time monitoring. Quantification curves were developed followed  by  testing  of  unknowns,  using  conventional  and  sigmoidal  model  fitting  (4  parametric)  approaches  whereas  both  peak  height  ratio  (PHR)  and  amplification  times  were  used  as  t hresholds.    Sensitivity  and  specificity  were  assessed  followed  by  testing  human  plasma  without  DNA  extraction.  A  positive  linear  relationship  of  peak  currents  of  SWV  vs  concentrations of MB was observed with significantly higher currents at higher temperat ure.  Incubation temperature of 63  o C and 25  o M of MB were the optimized condition for LAMP  based  on  reduction  of  PHRs.  Linear  relationships  between  logarithmic  concentrations  of  HBV DNA vs amplification time and PHRs depending on the thresholds were obtain ed with  different  coefficient  of  determination  (R 2 ) value.  Peak  height  thresholds  of  0.6  and  time  threshold of 22 min were found to be more suited in conventional approach while the former  varied to 0.7 and latter remined same in model fitting approach, ba sed on R 2 . Concentrations  of  the  unknowns  were  quantified  successfully  to  the  logarithmic  concentrations  of  DNA.  The a pproach was found to be specific for HBV as no PHR reduction for human genomic  DNA  and HBV DNA up to fg/ o l  was detected . Human plasma coul d be used directly as the  temp late after a pre - heat treatment of either  subsequent heating for 5 min at 95  o C and 100  o C or 125  o C for 10 min whereas the centrifugation step of the former increased the LAMP efficiency.</subfield>
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    <subfield code="a">Hepatitis B</subfield>
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    <subfield code="a">Loc Thai Nguyen,</subfield>
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    <subfield code="a">Anal, Anil Kumar,</subfield>
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    <subfield code="a">Sadiq, Muhammad Bilal,</subfield>
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    <subfield code="a">Asian Institute of Technology Fellowship,</subfield>
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    <subfield code="a">Asian Institute of Technology.</subfield>
    <subfield code="t">Thesis ;</subfield>
    <subfield code="v">no. no.FB-17-02</subfield>
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