日本感染症学雑誌第82巻第3号
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- うまじ うすい
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1 Mycoplasma peumoiae Mycoplasma peumoiae Mycoplasma peumoiae SDC M. peumoiae M. peumoiae M. geitalium Mycoplasma Mycoplasma peumoiae
2 M. peumoiae M. peumoiae M. peumoiae MycoplasmaM. geitalium M. homiis M. salvarium M. fermetas M. orale M. peumoiae M. geitalium Mycoplasma M. peumoiae Mycoplasma M. peumoiae SDC Hif Bst µ µ p M. peumoiae M. peumoiae M. peumoiaemycoplasma Mycoplasma µ M. peumoiaem. peumoiae M. peumoiae
3 Table Straistestedithisstudy Name Strai(Type) Referece Cotetoftemplate LAMP Tt(mi.) * FH ATCC Clicalisolate 25.0 Clicalisolate 25.8 M29 ATCC Mac ATCC FH NBRC440(= ATCC553) Mycoplasmahomiis ATCC234(Typestrai) Mycoplasmasalvarium ATCC23064(Typestrai) Mycoplasmafermetas ATCC9989(Typestrai) Mycoplasmaorale ATCC2374(Typestrai) Mycoplasmageitalum ATCC33530(Typestrai) Acietobacterbaumai ATCC9606(Typestrai) Acietobacterlwofi ATCC5309(Typestrai) Alcaligeesfaecalis ATCC9209 Achromobacterxylosoxidas ATCC2706(Typestrai) Chlamydiapeumoiae KKp Chlamydiapsitaci Bud Chlamydiatrachomatis L2/434/Bu Chlamydiatrachomatis D/UW3/Cx Chryseobacterium idologees ATCC29897(Typestrai) Coxielabureti Niemile Escherichiacoli ATCC775(Typestrai) Flavobacterium breve ATCC4339(Typestrai) Flavobacterium odoratum ATCC465(Typestrai) Haemophlusifluezae ATCC9795 Klebsielapeumoiae ATCC3883(Typestrai) Legioelapeumophla ATCC3352(Typestrai) Legioelalogbeachae ATCC33462(Typestrai) Legioelabozemai ATCC3327(Typestrai) Legioelamicdadei ATCC3328(Typestrai) Moraxelacatarhalis Pseudomoasaerugiosa ATCC27853 Pseudomoasfluoresces ATCC3525(Typestrai) Pseudomoasstutzeri ATCC7588(Typestrai) Seratiamarcesces ATCC800 Staphylococcusaureus ATCC25923 Staphylococcusepidermidis ATCC4990(Typestrai) Streptococcuspeumoiae ATCC4969 Streptococcuspyogees ATCC965 * Tt(mi)deotestheamplficatiotimerequiredforthethresholdvalue(0.07)ofthevelocityofturbidityicreaseo LA320C,Loopamprealtimeturbidimetertobeexceeded. µ µ µ µ
4 M. peumoiae Fig. NucleotidesequeceofM.peumoiaeSDCrepetitiveelemet(M35024)usedtodesigtheLAMP primers.dna sequecesusedfortheprimerdesigigareshowbythesolidarrows.therestrictiosite forhifi,locatedbetweefadb,isidicatedbythedotedbox.adtheoligoucleotidesequeceof thelampprimersform.peumoiaesdcdetectioareshowithesolidliebox. p M. peumoiae Hif M. peumoiae Mycoplasma
5 Fig.2 ProtocolsforDNA extractiofrom (A)respiratoryspecimesad(B)PPLO cultures. Table2 CompositiooftheLAMP reactiomix TrisHCl KCl (NH4)2SO4 MgSO4 Twee20 Betaie dntps FIP BIP F3 B3 LoopF LoopB BstDNA polymerase Volume 20mM (ph8.8) 0mM 0mM 8mM 0.% 0.8M.4mM each 40pmol 40pmol 0pmol 0pmol 20pmol 20pmol 8Uits 20μL
6 M. peumoiae Fig.3 ComparisoofsesitivitybetweeLAMP(A),6SrRNA estedpcr(b)adpestedpcr(c)usigserialdilutiosofm.peumoiaem29geomicdna.laesadsymbols:lae/ ;600,000,Lae2/ ;60,000,Lae3/ ;6,000, Lae4/ ;600,Lae5/ ;60,Lae6/ ;6,Lae7/ ;0.6,Lae8/ ;0(copies/test),adLaeM;00bpDNA ladder, LaeM2;λHid Idigest Fig.4 SesitivityofLAMPusigserialdilutiosofplas middna icludigthetargetregiofrom them.peu moiaem29geomicdna. Symbols: ;600,000, ;60,000, ;6,000, ;600, ;60, ;6, ;0.6, ;0(copies/test) Fig.5 Agarosegelelectrophoresisadrestrictioaaly sisofthelamp productsamplifiedfrom severalm. peumoiaestrais. Lae;NBRC440 (FH), Lae2;ATCC5492 (Mac), Lae3;ATCC29342 (M29),Lae4;ATCC553 (FH), Lae5adLae6;cliicalisolates,LaeM;00bpDNA ladder, Mycoplasma
7 Table3 ResultsofdetctioforM.peumoiaefrom clicalsamplesbylampadpcr. Respiratoryspecime sputum trachealaspirate pharygealswab pleuralfluid urie trasthoraciceedleaspirate PPLO culturefrom specime sputum trachealaspirate pharygealswab pleuralfluid urie trasthoraciceedleaspirate LAMP() (%) 24(.8%) 24(.8%) LAMP() (%) 9(22.%) 9(22.%) PCR * ( ) (%) 24(.8%) 24(.8%) PCR * ( ) (%) 9(22.%) 9(22.%) * stpcrofestedpcrtargetigthe6srrna gee M. salvarium M. orale M. peumoiae M. peumoiae M. peumoiae M. peumoiae p hmw crl M. peumoiae SDC p SDC M. peumoiae M. peumoiae M. peumoiae M. geitaliummycoplasma M. peumoiae p M. peumoiae SDC SDC SDC M. peumoiae
8 M. peumoiae Mycoplasma Mycoplasma Ureaplasma Mycoplasma peumoiae Mycoplasma peumoiae M. peumoiae Mycoplasma peumoiae Chlamydia peumoiae, Legioella peumophila Mycoplasma peumoiae et al. Salmoella Mycoplasma peumoiae et al. Mycoplasma geitalium
9 Mycoplasma peumoiae Mycoplasma peumoiae Mycoplasma peumoiae Mycoplasma peumoiae et al. Mycoplasma peumoiae Mycoplasma peumoiae Mycoplasma peumoiae Mycoplasma peumoiae SDC M. peumoiae M. peumoiae
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