%PDF-1.6
%
1 0 obj
<>
endobj
2 0 obj
<>stream
Wei Li, You-hong Fang, Hong-qiang Shen, De-hua Yang, Qiang Shu, Shi-qiang Shang
Evaluation of a real-time method of simultaneous amplification and testing in diagnosis of Mycoplasma pneumoniae infection in children with pneumonia
www.plosone.org
www.plosone.org
endstream
endobj
3 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 14 0 R/Annots 15 0 R/Contents 16 0 R/TrimBox[0 0 612 792]>>
endobj
15 0 obj
[17 0 R 18 0 R 19 0 R 20 0 R 21 0 R]
endobj
17 0 obj
<>/Border[0 0 0]/Dest(Rpone.0177842.ref012)>>
endobj
18 0 obj
<>/Border[0 0 0]/Dest(Rpone.0177842.ref014)>>
endobj
19 0 obj
<>/Border[0 0 0]/Dest(Rpone.0177842.ref015)>>
endobj
20 0 obj
<>/Border[0 0 0]/Dest(Rpone.0177842.ref016)>>
endobj
21 0 obj
<>/Border[0 0 0]/A 22 0 R>>
endobj
22 0 obj
<>
endobj
16 0 obj
[23 0 R 24 0 R 25 0 R 26 0 R 27 0 R 28 0 R 29 0 R 30 0 R 31 0 R 32 0 R 33 0 R]
endobj
23 0 obj
<>stream
q
0.83 0.64 0.02 0 k
357.2787 641.3102 m
366.7465 641.3102 l
h
f*
371.9622 641.3102 m
381.4299 641.3102 l
h
f*
465.789 541.0205 m
475.3134 541.0205 l
h
f*
234.7654 268.2142 m
244.2898 268.2142 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(real-time)Tj
3.855 0 Td
(PCR)Tj
2.0637 0 Td
(is)Tj
0.8447 0 Td
(incapable)Tj
4.0251 0 Td
(of)Tj
1.0148 0 Td
(distinguishing)Tj
5.9187 0 Td
(live)Tj
1.6101 0 Td
(bacteria)Tj
3.3789 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.4684 0 Td
(dead)Tj
2.1316 0 Td
(ones.)Tj
2.2847 0 Td
(Given)Tj
2.6306 0 Td
(the)Tj
1.474 0 Td
(draw-)Tj
-34.9056 -1.3039 Td
(backs)Tj
2.4377 0 Td
(of)Tj
1.0148 0 Td
(current)Tj
3.1861 0 Td
(tests,)Tj
2.1884 0 Td
(novel)Tj
2.3981 0 Td
(rapid)Tj
2.3414 0 Td
(diagnostic)Tj
4.32 0 Td
(techniques)Tj
4.524 0 Td
(with)Tj
2.0013 0 Td
(high)Tj
2.0069 0 Td
(sensitivity)Tj
4.2179 0 Td
(and)Tj
1.7348 0 Td
(specificity)Tj
-32.3714 -1.2982 Td
(are)Tj
1.4456 0 Td
(required)Tj
3.6397 0 Td
(for)Tj
1.3889 0 Td
(early)Tj
2.1487 0 Td
(identification)Tj
5.5842 0 Td
(of)Tj
/F7 1 Tf
1.0148 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F7 1 Tf
1.3209 0 Td
[()11()6()2()12()6()11()18()4( )6()]TJ
/F8 1 Tf
5.057 0 Td
(infections.)Tj
4.3767 0 Td
(Simultaneous)Tj
5.6579 0 Td
(amplification)Tj
-31.6344 -1.2983 Td
(and)Tj
1.7291 0 Td
(testing\(SAT\))Tj
5.3347 0 Td
(is)Tj
0.8448 0 Td
(a)Tj
0.6576 0 Td
(recently)Tj
3.4072 0 Td
(developed)Tj
4.269 0 Td
(method)Tj
3.3278 0 Td
(based)Tj
2.4718 0 Td
(on)Tj
1.2756 0 Td
(isothermal)Tj
4.473 0 Td
(amplification)Tj
5.5389 0 Td
(of)Tj
1.0148 0 Td
(RNA)Tj
-34.3443 -1.2983 Td
(and)Tj
1.7291 0 Td
(real-time)Tj
3.8551 0 Td
(detection)Tj
3.9344 0 Td
(of)Tj
1.0148 0 Td
(fluorescence)Tj
5.1874 0 Td
(which)Tj
2.6532 0 Td
(was)Tj
1.6951 0 Td
(previously)Tj
4.354 0 Td
(established)Tj
4.5978 0 Td
(to)Tj
1.0261 0 Td
(detect)Tj
/F7 1 Tf
2.6135 0 Td
[()47(\n)3()7()13()4( )6()-4(\r)11()]TJ
-32.6605 -1.3039 Td
[()-9()12()12()]TJ
2.1543 0 Td
[(\r)7()14()4()2()20()1()4()4()7()10()-4()]TJ
/F8 1 Tf
4.8018 0 Td
(and)Tj
1.7291 0 Td
(hepatitis)Tj
3.6114 0 Td
(C)Tj
0.8844 0 Td
(virus)Tj
2.2053 0 Td
([)Tj
0.83 0.64 0.02 0 k
(12)Tj
0 g
()Tj
0.83 0.64 0.02 0 k
(14)Tj
0 g
(].)Tj
3.5433 0 Td
(The)Tj
1.7801 0 Td
(aim)Tj
1.7405 0 Td
(of)Tj
1.0148 0 Td
(this)Tj
1.6781 0 Td
(study)Tj
2.3868 0 Td
(was)Tj
1.7007 0 Td
(to)Tj
1.0262 0 Td
(establish)Tj
3.6453 0 Td
(and)Tj
1.7291 0 Td
(eval-)Tj
-35.6312 -1.2982 Td
(uate)Tj
1.9048 0 Td
(potential)Tj
3.7587 0 Td
(utility)Tj
2.5795 0 Td
(of)Tj
1.0148 0 Td
(SAT)Tj
1.99 0 Td
(for)Tj
1.3889 0 Td
(early)Tj
2.1487 0 Td
(detection)Tj
3.9344 0 Td
(of)Tj
/F7 1 Tf
1.0148 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F7 1 Tf
1.321 0 Td
[()11()6()2()12()6()11()18()4( )6()]TJ
/F8 1 Tf
5.0569 0 Td
(in)Tj
1.0375 0 Td
(clinical)Tj
3.0728 0 Td
(practice.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 603.0992 cm
BT
/F3 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Methods)Tj
/F0 1 Tf
0 -1.4976 Td
(Patients)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 568.1196 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(From)Tj
2.4434 0 Td
(January)Tj
3.3222 0 Td
(2016)Tj
2.1203 0 Td
(to)Tj
1.0261 0 Td
(April)Tj
2.3074 0 Td
(2016,)Tj
2.3471 0 Td
(a)Tj
0.6519 0 Td
(total)Tj
2.007 0 Td
(of)Tj
1.0148 0 Td
(315)Tj
1.644 0 Td
(children)Tj
3.5547 0 Td
(were)Tj
2.1259 0 Td
(enrolled)Tj
3.515 0 Td
(in)Tj
1.0318 0 Td
(this)Tj
1.6781 0 Td
(study,)Tj
2.6135 0 Td
(including)Tj
-33.4032 -1.2982 Td
(202)Tj
1.644 0 Td
(boys)Tj
2.0523 0 Td
(and)Tj
1.7291 0 Td
(113)Tj
1.6441 0 Td
(girls.)Tj
2.16 0 Td
(The)Tj
1.7801 0 Td
(ages)Tj
1.9049 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(study)Tj
2.3925 0 Td
(participants)Tj
4.9549 0 Td
(ranged)Tj
2.9934 0 Td
(from)Tj
2.2053 0 Td
(1)Tj
0.6917 0 Td
(month)Tj
2.9253 0 Td
(to)Tj
1.0261 0 Td
(10)Tj
1.1679 0 Td
(years.)Tj
2.4888 0 Td
(All)Tj
-36.2435 -1.3039 Td
(study)Tj
2.3867 0 Td
(participants)Tj
4.9606 0 Td
(had)Tj
1.7065 0 Td
(been)Tj
2.1259 0 Td
(primarily)Tj
3.9685 0 Td
(diagnosed)Tj
4.2803 0 Td
(with)Tj
1.9956 0 Td
(pneumonia)Tj
4.8132 0 Td
([)Tj
0.83 0.64 0.02 0 k
(15)Tj
0 g
(])Tj
1.8538 0 Td
(and)Tj
1.7291 0 Td
(had)Tj
1.7121 0 Td
(received)Tj
3.532 0 Td
(no)Tj
-35.0643 -1.2983 Td
(clinical)Tj
3.0727 0 Td
(treatment.)Tj
4.3653 0 Td
(During)Tj
3.1351 0 Td
(study)Tj
2.3868 0 Td
(period,)Tj
3.0614 0 Td
(two)Tj
1.7064 0 Td
(respiratory)Tj
4.5978 0 Td
(tract)Tj
2.0466 0 Td
(samples)Tj
3.3845 0 Td
(\(throat)Tj
2.9934 0 Td
(swab)Tj
2.2053 0 Td
(or)Tj
1.0999 0 Td
(sputum\))Tj
-34.0552 -1.2983 Td
(from)Tj
2.2053 0 Td
(every)Tj
2.3471 0 Td
(child)Tj
2.211 0 Td
(were)Tj
2.1259 0 Td
(collected.)Tj
3.9629 0 Td
(One)Tj
1.9388 0 Td
(sample)Tj
3.0218 0 Td
(was)Tj
1.7007 0 Td
(mixed)Tj
2.7213 0 Td
(with)Tj
1.9955 0 Td
(1)Tj
0.6974 0 Td
(ml)Tj
1.2869 0 Td
(normal)Tj
3.1464 0 Td
(saline)Tj
2.5002 0 Td
(for)Tj
1.3833 0 Td
(real-time)Tj
-33.2445 -1.3039 Td
(PCR)Tj
2.0636 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4683 0 Td
(other)Tj
2.3471 0 Td
(was)Tj
1.7008 0 Td
(mixed)Tj
2.7212 0 Td
(with)Tj
2.0013 0 Td
(500)Tj
/F14 1 Tf
[()]TJ
/F8 1 Tf
(l)Tj
2.3981 0 Td
(RNA)Tj
2.262 0 Td
(protective)Tj
4.2066 0 Td
(agent)Tj
2.3867 0 Td
(\(RNase)Tj
3.1351 0 Td
(inhibitor)Tj
3.7814 0 Td
(and)Tj
1.7348 0 Td
(lysis)Tj
-33.9361 -1.2982 Td
(reagent\))Tj
3.5206 0 Td
(for)Tj
1.3889 0 Td
(SAT-MP.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 463.8614 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Ethics)Tj
2.9149 0 Td
(statement)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 446.8535 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(This)Tj
1.9842 0 Td
(study)Tj
2.3924 0 Td
(was)Tj
1.6951 0 Td
(approved)Tj
3.9742 0 Td
(by)Tj
1.1848 0 Td
(the)Tj
1.4684 0 Td
(ethics)Tj
2.5228 0 Td
(committee)Tj
4.4957 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(Children's)Tj
4.371 0 Td
(Hospital,)Tj
3.8438 0 Td
(Zhejiang)Tj
3.7417 0 Td
(Univer-)Tj
-34.1629 -1.3039 Td
(sity)Tj
1.5987 0 Td
(School)Tj
2.9083 0 Td
(of)Tj
1.0148 0 Td
(Medicine,)Tj
4.2179 0 Td
(and)Tj
1.7292 0 Td
(written)Tj
3.1124 0 Td
(consent)Tj
3.3335 0 Td
(was)Tj
1.6951 0 Td
(obtained)Tj
3.736 0 Td
(from)Tj
2.2054 0 Td
(patients')Tj
3.583 0 Td
(parents)Tj
3.1747 0 Td
(or)Tj
1.0942 0 Td
(legal)Tj
-33.4032 -1.2982 Td
(guardians.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 394.5826 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Serological)Tj
5.0929 0 Td
(test)Tj
1.8236 0 Td
(for)Tj
1.3749 0 Td
(detection)Tj
4.2283 0 Td
(of)Tj
1.0441 0 Td
(IgM)Tj
1.8756 0 Td
(antibody)Tj
3.9497 0 Td
(of)Tj
/F6 1 Tf
1.0488 0 Td
(M)Tj
/F0 1 Tf
(.)Tj
/F6 1 Tf
1.3229 0 Td
(pneumoniae)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 377.5748 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Blood)Tj
2.5851 0 Td
(samples)Tj
3.3846 0 Td
(and)Tj
1.7291 0 Td
(respiratory)Tj
4.5978 0 Td
(tract)Tj
2.0466 0 Td
(samples)Tj
3.3788 0 Td
(were)Tj
2.126 0 Td
(collected)Tj
3.7361 0 Td
(at)Tj
0.9524 0 Td
(the)Tj
1.474 0 Td
(same)Tj
2.2564 0 Td
(time.)Tj
2.2507 0 Td
(The)Tj
1.7858 0 Td
(serum)Tj
2.7212 0 Td
(was)Tj
-35.0246 -1.3039 Td
(separated)Tj
4.0081 0 Td
(and)Tj
1.7291 0 Td
(then)Tj
2.0183 0 Td
(stored)Tj
2.7099 0 Td
(at)Tj
0.9524 0 Td
(-70C)Tj
2.5795 0 Td
(till)Tj
1.2756 0 Td
(the)Tj
1.4684 0 Td
(time)Tj
2.0296 0 Td
(of)Tj
1.0148 0 Td
(analysis.)Tj
3.5546 0 Td
(The)Tj
/F7 1 Tf
1.7801 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F7 1 Tf
1.3153 0 Td
[()11()6()2()12()6()11()18()4( )6()]TJ
/F8 1 Tf
5.0626 0 Td
(IgM)Tj
1.9162 0 Td
(was)Tj
1.7008 0 Td
(deter-)Tj
-35.1153 -1.2982 Td
(mined)Tj
2.8062 0 Td
(via)Tj
1.3777 0 Td
(a)Tj
0.6519 0 Td
(specific)Tj
3.1918 0 Td
(Anti-)Tj
/F7 1 Tf
[()]TJ
/F8 1 Tf
(.)Tj
/F7 1 Tf
3.4639 0 Td
[()11()6()2()12()6()11()18()4( )6()]TJ
/F8 1 Tf
5.057 0 Td
(ELISA)Tj
2.829 0 Td
(assay)Tj
2.2677 0 Td
(\(EUROIMMUN,)Tj
7.0582 0 Td
(Luebeck,)Tj
3.7814 0 Td
(Germany\),)Tj
-32.4848 -1.2983 Td
(the)Tj
1.4683 0 Td
(absorbance)Tj
4.7338 0 Td
(above)Tj
2.5512 0 Td
(1.1)Tj
1.3946 0 Td
(was)Tj
1.7008 0 Td
(determined)Tj
4.8472 0 Td
(positive.)Tj
3.5433 0 Td
(All)Tj
1.3946 0 Td
(performance)Tj
5.3518 0 Td
(was)Tj
1.6951 0 Td
(according)Tj
4.1896 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(man-)Tj
-35.3648 -1.2983 Td
(ufacturer's)Tj
4.4617 0 Td
(instructions.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 299.2818 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Real-time)Tj
4.4457 0 Td
(PCR)Tj
2.3102 0 Td
(for)Tj
1.3795 0 Td
(detection)Tj
4.2284 0 Td
(of)Tj
/F6 1 Tf
1.0441 0 Td
(M)Tj
/F0 1 Tf
(.)Tj
/F6 1 Tf
1.3276 0 Td
(pneumoniae)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 282.274 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(primers)Tj
3.3505 0 Td
(and)Tj
1.7348 0 Td
(probe)Tj
2.5455 0 Td
(were)Tj
2.1203 0 Td
(reported)Tj
3.6624 0 Td
(in)Tj
1.0318 0 Td
(previous)Tj
3.651 0 Td
(study)Tj
2.3867 0 Td
(which)Tj
2.6532 0 Td
(target)Tj
2.5115 0 Td
(the)Tj
1.4683 0 Td
(p1)Tj
1.2133 0 Td
(gene)Tj
2.0806 0 Td
(of)Tj
/F7 1 Tf
1.0148 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F7 1 Tf
1.3152 0 Td
[()11()6()2()]TJ
-34.52 -1.2982 Td
[()6()11()18()4( )6()]TJ
/F8 1 Tf
3.135 0 Td
([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
(].)Tj
2.075 0 Td
(1)Tj
0.6973 0 Td
(ml)Tj
1.2869 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(mixture)Tj
3.3959 0 Td
(containing)Tj
4.524 0 Td
(sample)Tj
3.0217 0 Td
(and)Tj
1.7292 0 Td
(normal)Tj
3.1464 0 Td
(saline)Tj
2.5001 0 Td
(was)Tj
1.7008 0 Td
(moved)Tj
2.9537 0 Td
(into)Tj
1.8425 0 Td
(a)Tj
-34.4917 -1.3039 Td
(1.5-ml)Tj
2.8119 0 Td
(microcentrifuge)Tj
6.6444 0 Td
(tube)Tj
1.9785 0 Td
(aseptically)Tj
4.3257 0 Td
(and)Tj
1.7291 0 Td
(centrifuged)Tj
4.7849 0 Td
(for)Tj
1.3833 0 Td
(5)Tj
0.6973 0 Td
(min.)Tj
2.0806 0 Td
(The)Tj
1.7858 0 Td
(supernatant)Tj
4.9776 0 Td
(was)Tj
-33.1991 -1.2983 Td
(decanted)Tj
3.8267 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4684 0 Td
(cell)Tj
1.559 0 Td
(pellets)Tj
2.7383 0 Td
(were)Tj
2.1203 0 Td
(suspended)Tj
4.439 0 Td
(in)Tj
1.0375 0 Td
(50)Tj
/F14 1 Tf
[()]TJ
/F8 1 Tf
(l)Tj
1.9218 0 Td
(of)Tj
1.0148 0 Td
(lysis)Tj
1.9106 0 Td
(buffer)Tj
2.6248 0 Td
(\(Da'an)Tj
2.931 0 Td
(gene)Tj
2.0807 0 Td
(company,)Tj
-31.402 -1.2983 Td
(China\).)Tj
3.2258 0 Td
(And)Tj
1.9785 0 Td
(the)Tj
1.4684 0 Td
(mixture)Tj
3.3958 0 Td
(was)Tj
1.7008 0 Td
(then)Tj
2.0183 0 Td
(boiled)Tj
2.6985 0 Td
(for)Tj
1.3833 0 Td
(10)Tj
1.1736 0 Td
(min.)Tj
2.0806 0 Td
(4)Tj
/F14 1 Tf
[()]TJ
/F8 1 Tf
(l)Tj
1.4457 0 Td
(of)Tj
1.0148 0 Td
(supernatant)Tj
4.9832 0 Td
(was)Tj
1.7008 0 Td
(served)Tj
2.7779 0 Td
(as)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(tem-)Tj
-35.5348 -1.2982 Td
(plate)Tj
2.1373 0 Td
(in)Tj
1.0374 0 Td
(real-time)Tj
3.8551 0 Td
(PCR)Tj
2.0636 0 Td
(system,)Tj
3.1635 0 Td
(containing)Tj
4.524 0 Td
(400)Tj
1.6498 0 Td
(nM)Tj
1.6611 0 Td
(of)Tj
1.0148 0 Td
(each)Tj
2.0296 0 Td
(forward)Tj
3.4015 0 Td
(and)Tj
1.7292 0 Td
(reverse)Tj
3.05 0 Td
(primer,)Tj
3.2145 0 Td
(100)Tj
-34.5314 -1.3039 Td
(nM)Tj
1.661 0 Td
(of)Tj
1.0148 0 Td
(fluorescently-labeled)Tj
8.4983 0 Td
(specific)Tj
3.1917 0 Td
(probe\(5'HEX3'BHQ1\),)Tj
10.3124 0 Td
(and)Tj
1.7348 0 Td
(25)Tj
/F14 1 Tf
[()]TJ
/F8 1 Tf
(l)Tj
1.9219 0 Td
(of)Tj
1.0148 0 Td
(Taqman)Tj
3.5716 0 Td
(master)Tj
2.931 0 Td
(mix)Tj
-35.8523 -1.2983 Td
(\(2)Tj
/F14 1 Tf
[()]TJ
/F8 1 Tf
(,)Tj
1.8311 0 Td
(Vazym,)Tj
3.2598 0 Td
(China\))Tj
2.9991 0 Td
(with)Tj
2.0012 0 Td
(water)Tj
2.4265 0 Td
(added)Tj
2.6645 0 Td
(to)Tj
1.0262 0 Td
(give)Tj
1.8255 0 Td
(a)Tj
0.6576 0 Td
(final)Tj
2.0012 0 Td
(volume)Tj
3.1975 0 Td
(of)Tj
1.0148 0 Td
(50)Tj
/F14 1 Tf
[()]TJ
/F8 1 Tf
(l)Tj
1.9275 0 Td
(per)Tj
1.5307 0 Td
(sample.)Tj
3.2485 0 Td
(P)Tj
0.7767 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(real-)Tj
-34.8942 -1.2983 Td
(time)Tj
2.0295 0 Td
(PCR)Tj
2.0637 0 Td
(amplification)Tj
5.5331 0 Td
(performed)Tj
4.4731 0 Td
(on)Tj
1.2756 0 Td
(an)Tj
1.2019 0 Td
(ABI)Tj
1.8255 0 Td
(7500)Tj
2.1203 0 Td
(detection)Tj
3.9401 0 Td
(system)Tj
2.9367 0 Td
(in)Tj
1.0374 0 Td
(following)Tj
3.9742 0 Td
(conditions:)Tj
-32.4111 -1.3039 Td
(94C)Tj
2.228 0 Td
(for)Tj
1.3833 0 Td
(2)Tj
0.6973 0 Td
(min)Tj
1.8538 0 Td
(and)Tj
1.7292 0 Td
(40)Tj
1.1735 0 Td
(cycles)Tj
2.5512 0 Td
(of)Tj
1.0148 0 Td
(94C)Tj
2.228 0 Td
(for)Tj
1.3833 0 Td
(15)Tj
1.1735 0 Td
(s)Tj
0.5783 0 Td
(and)Tj
1.7348 0 Td
(60C)Tj
2.2223 0 Td
(for)Tj
1.389 0 Td
(45s.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 139.011 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(SAT-MP)Tj
3.9779 0 Td
(for)Tj
1.3748 0 Td
(detection)Tj
4.2284 0 Td
(of)Tj
/F6 1 Tf
1.0441 0 Td
(M)Tj
/F0 1 Tf
(.)Tj
/F6 1 Tf
1.3276 0 Td
(pneumoniae)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 122.0031 cm
BT
/F8 1 Tf
1 TL
0.01 Tc
0 0 Td
(In)Tj
1.1338 0 Td
(the)Tj
1.5137 0 Td
(SAT)Tj
2.0296 0 Td
(technology,)Tj
5.0059 0 Td
(16S)Tj
1.6838 0 Td
(rRNA)Tj
2.6929 0 Td
(was)Tj
1.7405 0 Td
(transcribed)Tj
4.9039 0 Td
(into)Tj
1.8992 0 Td
(cDNA)Tj
2.8573 0 Td
(equipped)Tj
4.0422 0 Td
(with)Tj
2.0522 0 Td
(the)Tj
1.5137 0 Td
(T7)Tj
1.3323 0 Td
(pro-)Tj
-34.401 -1.2982 Td
(moter)Tj
2.7099 0 Td
(with)Tj
2.0522 0 Td
(the)Tj
1.5137 0 Td
(help)Tj
1.9899 0 Td
(of)Tj
1.0432 0 Td
(Moloney)Tj
3.8947 0 Td
(murine)Tj
3.2599 0 Td
(leukemia)Tj
3.9741 0 Td
(virus)Tj
2.2677 0 Td
(\(MLV\))Tj
3.0954 0 Td
(reverse)Tj
3.1465 0 Td
(transcriptase)Tj
5.5162 0 Td
(and)Tj
-34.4634 -1.3039 Td
(then)Tj
2.0749 0 Td
(multiple)Tj
3.6737 0 Td
(RNA)Tj
2.3074 0 Td
(copies\(100)Tj
4.6261 0 Td
(~)Tj
0.8107 0 Td
(1000\))Tj
2.5341 0 Td
(were)Tj
2.177 0 Td
(produced)Tj
4.1613 0 Td
(from)Tj
2.262 0 Td
(each)Tj
2.0806 0 Td
(cDNA)Tj
2.8573 0 Td
(copy)Tj
2.177 0 Td
(by)Tj
1.2132 0 Td
(use)Tj
1.5704 0 Td
(of)Tj
1.0432 0 Td
(T7)Tj
-35.5689 -1.2983 Td
(RNA)Tj
2.3017 0 Td
(polymerase.)Tj
5.159 0 Td
(Afterwards,)Tj
5.04 0 Td
(these)Tj
2.3244 0 Td
(RNA)Tj
2.3073 0 Td
(copies)Tj
2.8007 0 Td
(were)Tj
2.1769 0 Td
(transcribed)Tj
4.9096 0 Td
(into)Tj
1.8936 0 Td
(cDNAs)Tj
3.2314 0 Td
(again)Tj
2.4378 0 Td
(and)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 107.1496 23.6976 re
W* n
q
107.0929 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 363.4582 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(SAT-MP)Tj
3.9614 0 Td
(in)Tj
0.978 0 Td
(diagnos)Tj
3.4299 0 Td
(is)Tj
0.9284 0 Td
(of)Tj
/F6 1 Tf
1.0417 0 Td
(Mycoplas)Tj
4.1528 0 Td
(ma)Tj
1.5945 0 Td
(pneumon)Tj
4.0961 0 Td
(iae)Tj
/F0 1 Tf
1.5236 0 Td
(pneumon)Tj
4.0961 0 Td
(ia)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01778)Tj
4.089 0 Td
(42)Tj
0 g
2.0906 0 Td
(May)Tj
2.0906 0 Td
(16,)Tj
1.5803 0 Td
(2017)Tj
35.7806 0 Td
(2)Tj
0.7654 0 Td
(/)Tj
0.496 0 Td
(7)Tj
/F10 1 Tf
-0.025 Tc
-66.9477 84.0904 Td
(Competing)Tj
4.3795 0 Td
(interests)Tj
3.3307 0 Td
(:)Tj
/F11 1 Tf
0.4819 0 Td
(The)Tj
1.5875 0 Td
(authors)Tj
3.0401 0 Td
(have)Tj
1.963 0 Td
(declared)Tj
-14.7827 -1.3748 Td
(that)Tj
1.6228 0 Td
(no)Tj
1.1693 0 Td
(competing)Tj
4.1599 0 Td
(interests)Tj
ET
endstream
endobj
24 0 obj
<>stream
endstream
endobj
25 0 obj
<>stream
endstream
endobj
26 0 obj
<>stream
endstream
endobj
27 0 obj
<>stream
endstream
endobj
28 0 obj
<>stream
endstream
endobj
29 0 obj
<>stream
endstream
endobj
30 0 obj
<>stream
endstream
endobj
31 0 obj
<>stream
endstream
endobj
32 0 obj
<>stream
endstream
endobj
33 0 obj
<>stream
BT
10.3394 82.7156 Td
(exist.)Tj
ET
Q
endstream
endobj
34 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 14 0 R/Annots 37 0 R/Contents 38 0 R/TrimBox[0 0 612 792]>>
endobj
35 0 obj
<>
endobj
39 0 obj
<>stream
/CIDInit/ProcSet findresource begin
12 dict begin begincmap
/CMapName/Adobe-Identity-UCS def
/CIDSystemInfo<>def
/CMapType 2 def
1 begincodespacerange
<0000>
endcodespacerange
18 beginbfchar
<0010><002D>
<0016><0033>
<0018><0035>
<0024><0041>
<0025><0042>
<0026><0043>
<0027><0044>
<0028><0045>
<0029><0046>
<002A><0047>
<002B><0048>
<002F><004C>
<0030><004D>
<0037><0054>
<0038><0055>
<003B><0058>
<003C><0059>
<00B6><2019>
endbfchar
endcmap
CMapName currentdict /CMap defineresource pop end end
endstream
endobj
40 0 obj
<>/FontDescriptor 41 0 R/DW 600>>
endobj
41 0 obj
<>
endobj
42 0 obj
<>stream
`DSIG EBDTL\ iX EBLC+\ mL 4GDEF̥
r GSUBLcu| s NOS/2
0M VVDMX2M' B bdatL\ bloc+\ 4cmapX[ Bcvt a d fpgm62 _ gasp + @ glyf v head)x l 6hhea{<