               SEQUENCE LISTING

<110> Universitätsklinikum Bonn (UKB)

<120> RT-LAMP sensitivity increase using unpurified biosamples

<130> AD3527 PCT

<150> EP 21 178 783.3
<151> 2021-06-10

<160> 40

<170> BiSSAP 1.3.6

<210> 1
<211> 592
<212> PRT
<213> Geobacillus stearothermophilus


<220> 
<223> Geobacillus strain

<400> 1
Met Glu Ser Pro Ser Ser Glu Glu Glu Lys Pro Leu Ala Lys Met Ala 
1               5                   10                  15      
Phe Thr Leu Ala Asp Arg Val Thr Glu Glu Met Leu Ala Asp Lys Ala 
            20                  25                  30          
Ala Leu Val Val Glu Val Val Glu Glu Asn Tyr His Asp Ala Pro Ile 
        35                  40                  45              
Val Gly Ile Ala Val Val Asn Glu His Gly Arg Phe Phe Leu Arg Pro 
    50                  55                  60                  
Glu Thr Ala Leu Ala Asp Pro Gln Phe Val Ala Trp Leu Gly Asp Glu 
65                  70                  75                  80  
Thr Lys Lys Lys Ser Met Phe Asp Ser Lys Arg Ala Ala Val Ala Leu 
                85                  90                  95      
Lys Trp Lys Gly Ile Glu Leu Cys Gly Val Ser Phe Asp Leu Leu Leu 
            100                 105                 110         
Ala Ala Tyr Leu Leu Asp Pro Ala Gln Gly Val Asp Asp Val Ala Ala 
        115                 120                 125             
Ala Ala Lys Met Lys Gln Tyr Glu Ala Val Arg Pro Asp Glu Ala Val 
    130                 135                 140                 
Tyr Gly Lys Gly Ala Lys Arg Ala Val Pro Asp Glu Pro Val Leu Ala 
145                 150                 155                 160 
Glu His Leu Val Arg Lys Ala Ala Ala Ile Trp Glu Leu Glu Arg Pro 
                165                 170                 175     
Phe Leu Asp Glu Leu Arg Arg Asn Glu Gln Asp Arg Leu Leu Val Glu 
            180                 185                 190         
Leu Glu Gln Pro Leu Ser Ser Ile Leu Ala Glu Met Glu Phe Ala Gly 
        195                 200                 205             
Val Lys Val Asp Thr Lys Arg Leu Glu Gln Met Gly Lys Glu Leu Ala 
    210                 215                 220                 
Glu Gln Leu Gly Thr Val Glu Gln Arg Ile Tyr Glu Leu Ala Gly Gln 
225                 230                 235                 240 
Glu Phe Asn Ile Asn Ser Pro Lys Gln Leu Gly Val Ile Leu Phe Glu 
                245                 250                 255     
Lys Leu Gln Leu Pro Val Leu Lys Lys Thr Lys Thr Gly Tyr Ser Thr 
            260                 265                 270         
Ser Ala Asp Val Leu Glu Lys Leu Ala Pro Tyr His Glu Ile Val Glu 
        275                 280                 285             
Asn Ile Leu His Tyr Arg Gln Leu Gly Lys Leu Gln Ser Thr Tyr Ile 
    290                 295                 300                 
Glu Gly Leu Leu Lys Val Val Arg Pro Asp Thr Lys Lys Val His Thr 
305                 310                 315                 320 
Ile Phe Asn Gln Ala Leu Thr Gln Thr Gly Arg Leu Ser Ser Thr Glu 
                325                 330                 335     
Pro Asn Leu Gln Asn Ile Pro Ile Arg Leu Glu Glu Gly Arg Lys Ile 
            340                 345                 350         
Arg Gln Ala Phe Val Pro Ser Glu Ser Asp Trp Leu Ile Phe Ala Ala 
        355                 360                 365             
Asp Tyr Ser Gln Ile Glu Leu Arg Val Leu Ala His Ile Ala Glu Asp 
    370                 375                 380                 
Asp Asn Leu Met Glu Ala Phe Arg Arg Asp Leu Asp Ile His Thr Lys 
385                 390                 395                 400 
Thr Ala Met Asp Ile Phe Gln Val Ser Glu Asp Glu Val Thr Pro Asn 
                405                 410                 415     
Met Arg Arg Gln Ala Lys Ala Val Asn Phe Gly Ile Val Tyr Gly Ile 
            420                 425                 430         
Ser Asp Tyr Gly Leu Ala Gln Asn Leu Asn Ile Ser Arg Lys Glu Ala 
        435                 440                 445             
Ala Glu Phe Ile Glu Arg Tyr Phe Glu Ser Phe Pro Gly Val Lys Arg 
    450                 455                 460                 
Tyr Met Glu Asn Ile Val Gln Glu Ala Lys Gln Lys Gly Tyr Val Thr 
465                 470                 475                 480 
Thr Leu Leu His Arg Arg Arg Tyr Leu Pro Asp Ile Thr Ser Arg Asn 
                485                 490                 495     
Phe Asn Val Arg Ser Phe Ala Glu Arg Met Ala Met Asn Thr Pro Ile 
            500                 505                 510         
Gln Gly Ser Ala Ala Asp Ile Ile Lys Lys Ala Met Ile Asp Leu Asn 
        515                 520                 525             
Ala Arg Leu Lys Glu Glu Arg Leu Gln Ala His Leu Leu Leu Gln Val 
    530                 535                 540                 
His Asp Glu Leu Ile Leu Glu Ala Pro Lys Glu Glu Met Glu Arg Leu 
545                 550                 555                 560 
Cys Arg Leu Val Pro Glu Val Met Glu Gln Ala Val Thr Leu Arg Val 
                565                 570                 575     
Pro Leu Lys Val Asp Tyr His Tyr Gly Ser Thr Trp Tyr Asp Ala Lys 
            580                 585                 590         



<210> 2
<211> 26
<212> DNA
<213> Artificial Sequence


<220> 
<223> E_Sarbeco_F1

<400> 2
acaggtacgt taatagttaa tagcgt                                         26


<210> 3
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> E_Sarbeco_R2

<400> 3
atattgcagc agtacgcaca ca                                             22


<210> 4
<211> 26
<212> DNA
<213> Artificial Sequence


<220> 
<223> E_Sarbeco_P1

<400> 4
acactagcca tccttactgc gcttcg                                         26


<210> 5
<211> 16
<212> DNA
<213> Artificial Sequence


<220> 
<223> MS2_F

<400> 5
tgctcgcgga tacccg                                                    16


<210> 6
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> MS2_R

<400> 6
aacttgcgtt ctcgagcgat                                                20


<210> 7
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> MS2_P

<400> 7
acctcgggtt tccgtcttgc tcgt                                           24


<210> 8
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> RT Primer

<400> 8
gtaagaacac ctgtgcctgt ta                                             22


<210> 9
<211> 55
<212> DNA
<213> Artificial Sequence


<220> 
<223> Forward Primer

<400> 9
acactctttc cctacacgac gctcttccga tctagacttt ttaggtccac aaaca          55


<210> 10
<211> 56
<212> DNA
<213> Artificial Sequence


<220> 
<223> Reverse Primer

<400> 10
tgactggagt tcagacgtgt gctcttccga tctccggtag cacaccttgt aatggt         56


<210> 11
<211> 51
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-FIP-BC

<400> 11
tgcggccaat gtttgtaatc agagcttaca ctccaaggaa attttgggga c              51


<210> 12
<211> 39
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-BIP

<400> 12
cgcattggca tggaagtcac tttgatggca cctgtgtag                            39


<210> 13
<211> 19
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-LF

<400> 13
ttccttgtct gattagttc                                                 19


<210> 14
<211> 18
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-LB

<400> 14
accttcggga acgtggtt                                                  18


<210> 15
<211> 18
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-F3

<400> 15
aacacaagct ttcggcag                                                  18


<210> 16
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-B3

<400> 16
gaaatttgga tctttgtcat cc                                             22


<210> 17
<211> 18
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-F3-LNA

<400> 17
aacacaagct ttcggcag                                                  18


<210> 18
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-B3-LNA

<400> 18
gaaatttgga tctttgtcat cc                                             22


<210> 19
<211> 56
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-PCR forward

<400> 19
acactctttc cctacacgac gctcttccga tctaacgctg aagcgctggg ggcaaa         56


<210> 20
<211> 56
<212> DNA
<213> Artificial Sequence


<220> 
<223> SARS-CoV-2-PCR reverse

<400> 20
tgactggagt tcagacgtgt gctcttccga tctgtttgta atcagttcct tgtctg         56


<210> 21
<211> 49
<212> DNA
<213> Artificial Sequence


<220> 
<223> Influenza-A-FIP

<220> 
<221> variation
<222> 23
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 24
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 25
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 26
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 27
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 28
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 29
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 30
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 31
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 32
<223> /replace="a,c,g,t"

<400> 21
ttagtcagag gtgacarrat tgnnnnnnnn nncagatctt gaggctctc                 49


<210> 22
<211> 39
<212> DNA
<213> Artificial Sequence


<220> 
<223> Influenza-A-BIP

<400> 22
ttgtkttcac gctcaccgtg tttggacaaa gcgtctacg                            39


<210> 23
<211> 16
<212> DNA
<213> Artificial Sequence


<220> 
<223> Influenza-A-LF

<400> 23
gtcttgtctt tagcca                                                    16


<210> 24
<211> 17
<212> DNA
<213> Artificial Sequence


<220> 
<223> Influenza-A-LB

<400> 24
cmagtgagcg aggactg                                                   17


<210> 25
<211> 19
<212> DNA
<213> Artificial Sequence


<220> 
<223> Influenza-A-F3-1 5x LNA

<400> 25
gacttgaaga tgtctttgc                                                 19


<210> 26
<211> 19
<212> DNA
<213> Artificial Sequence


<220> 
<223> Influenza-A-F3-2 5x LNA

<400> 26
gactggaaag tgtctttgc                                                 19


<210> 27
<211> 19
<212> DNA
<213> Artificial Sequence


<220> 
<223> Influenza-A-B3-1 5x LNA

<400> 27
trttatttgg gtctccatt                                                 19


<210> 28
<211> 19
<212> DNA
<213> Artificial Sequence


<220> 
<223> Influenza-A-B3-2 5x LNA

<400> 28
trttgtttgg gtccccatt                                                 19


<210> 29
<211> 159
<212> DNA
<213> SARS-CoV-2


<220> 
<223> SARS-CoV-2 spike gene

<400> 29
agacttttta ggtccacaaa cagttgctgg tgcatgtaga agttcaaaag aaagtactac     60

tactctgtta tggtttggta accaacacca taagtgtggt ttggaaacca tatgtattgt    120

taaaggaaag taacaattaa aacccttcaa ccacccatt                           159


<210> 30
<211> 611
<212> PRT
<213> Geobacillus stearothermophilus


<220> 
<223> Geobacillus strain

<400> 30
Met Lys His His His His His His Ser Ala Gly Leu Glu Val Leu Phe 
1               5                   10                  15      
Gln Gly Pro Met Glu Ser Pro Ser Ser Glu Glu Glu Lys Pro Leu Ala 
            20                  25                  30          
Lys Met Ala Phe Thr Leu Ala Asp Arg Val Thr Glu Glu Met Leu Ala 
        35                  40                  45              
Asp Lys Ala Ala Leu Val Val Glu Val Val Glu Glu Asn Tyr His Asp 
    50                  55                  60                  
Ala Pro Ile Val Gly Ile Ala Val Val Asn Glu His Gly Arg Phe Phe 
65                  70                  75                  80  
Leu Arg Pro Glu Thr Ala Leu Ala Asp Pro Gln Phe Val Ala Trp Leu 
                85                  90                  95      
Gly Asp Glu Thr Lys Lys Lys Ser Met Phe Asp Ser Lys Arg Ala Ala 
            100                 105                 110         
Val Ala Leu Lys Trp Lys Gly Ile Glu Leu Cys Gly Val Ser Phe Asp 
        115                 120                 125             
Leu Leu Leu Ala Ala Tyr Leu Leu Asp Pro Ala Gln Gly Val Asp Asp 
    130                 135                 140                 
Val Ala Ala Ala Ala Lys Met Lys Gln Tyr Glu Ala Val Arg Pro Asp 
145                 150                 155                 160 
Glu Ala Val Tyr Gly Lys Gly Ala Lys Arg Ala Val Pro Asp Glu Pro 
                165                 170                 175     
Val Leu Ala Glu His Leu Val Arg Lys Ala Ala Ala Ile Trp Glu Leu 
            180                 185                 190         
Glu Arg Pro Phe Leu Asp Glu Leu Arg Arg Asn Glu Gln Asp Arg Leu 
        195                 200                 205             
Leu Val Glu Leu Glu Gln Pro Leu Ser Ser Ile Leu Ala Glu Met Glu 
    210                 215                 220                 
Phe Ala Gly Val Lys Val Asp Thr Lys Arg Leu Glu Gln Met Gly Lys 
225                 230                 235                 240 
Glu Leu Ala Glu Gln Leu Gly Thr Val Glu Gln Arg Ile Tyr Glu Leu 
                245                 250                 255     
Ala Gly Gln Glu Phe Asn Ile Asn Ser Pro Lys Gln Leu Gly Val Ile 
            260                 265                 270         
Leu Phe Glu Lys Leu Gln Leu Pro Val Leu Lys Lys Thr Lys Thr Gly 
        275                 280                 285             
Tyr Ser Thr Ser Ala Asp Val Leu Glu Lys Leu Ala Pro Tyr His Glu 
    290                 295                 300                 
Ile Val Glu Asn Ile Leu His Tyr Arg Gln Leu Gly Lys Leu Gln Ser 
305                 310                 315                 320 
Thr Tyr Ile Glu Gly Leu Leu Lys Val Val Arg Pro Asp Thr Lys Lys 
                325                 330                 335     
Val His Thr Ile Phe Asn Gln Ala Leu Thr Gln Thr Gly Arg Leu Ser 
            340                 345                 350         
Ser Thr Glu Pro Asn Leu Gln Asn Ile Pro Ile Arg Leu Glu Glu Gly 
        355                 360                 365             
Arg Lys Ile Arg Gln Ala Phe Val Pro Ser Glu Ser Asp Trp Leu Ile 
    370                 375                 380                 
Phe Ala Ala Asp Tyr Ser Gln Ile Glu Leu Arg Val Leu Ala His Ile 
385                 390                 395                 400 
Ala Glu Asp Asp Asn Leu Met Glu Ala Phe Arg Arg Asp Leu Asp Ile 
                405                 410                 415     
His Thr Lys Thr Ala Met Asp Ile Phe Gln Val Ser Glu Asp Glu Val 
            420                 425                 430         
Thr Pro Asn Met Arg Arg Gln Ala Lys Ala Val Asn Phe Gly Ile Val 
        435                 440                 445             
Tyr Gly Ile Ser Asp Tyr Gly Leu Ala Gln Asn Leu Asn Ile Ser Arg 
    450                 455                 460                 
Lys Glu Ala Ala Glu Phe Ile Glu Arg Tyr Phe Glu Ser Phe Pro Gly 
465                 470                 475                 480 
Val Lys Arg Tyr Met Glu Asn Ile Val Gln Glu Ala Lys Gln Lys Gly 
                485                 490                 495     
Tyr Val Thr Thr Leu Leu His Arg Arg Arg Tyr Leu Pro Asp Ile Thr 
            500                 505                 510         
Ser Arg Asn Phe Asn Val Arg Ser Phe Ala Glu Arg Met Ala Met Asn 
        515                 520                 525             
Thr Pro Ile Gln Gly Ser Ala Ala Asp Ile Ile Lys Lys Ala Met Ile 
    530                 535                 540                 
Asp Leu Asn Ala Arg Leu Lys Glu Glu Arg Leu Gln Ala His Leu Leu 
545                 550                 555                 560 
Leu Gln Val His Asp Glu Leu Ile Leu Glu Ala Pro Lys Glu Glu Met 
                565                 570                 575     
Glu Arg Leu Cys Arg Leu Val Pro Glu Val Met Glu Gln Ala Val Thr 
            580                 585                 590         
Leu Arg Val Pro Leu Lys Val Asp Tyr His Tyr Gly Ser Thr Trp Tyr 
        595                 600                 605             
Asp Ala Lys 
    610     

<210> 31
<211> 51
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-FIP-BC

<220> 
<221> variation
<222> 23
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 24
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 25
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 26
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 27
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 28
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 29
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 30
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 31
<223> /replace="a,c,g,t"

<220> 
<221> variation
<222> 32
<223> /replace="a,c,g,t"

<400> 31
gctgcctgga gttgaatttc ttnnnnnnnn nncaagcctc ttctcgttcc t              51


<210> 32
<211> 39
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-BIP

<400> 32
ctgctcttgc tttgctgctg taccagacat tttgctctc                            39


<210> 33
<211> 19
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-LF

<400> 33
gaactgttgc gactacgtg                                                 19


<210> 34
<211> 18
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-LB

<400> 34
cttgacagat tgaaccag                                                  18


<210> 35
<211> 18
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-F3 LNA

<400> 35
aaaggcttct acgcagaa                                                  18


<210> 36
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-B3 LNA

<400> 36
tttggccttg ttgttgttgg cc                                             22


<210> 37
<211> 18
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-F3 no LNA

<400> 37
aaaggcttct acgcagaa                                                  18


<210> 38
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-B3 no LNA

<400> 38
tttggccttg ttgttgttgg cc                                             22


<210> 39
<211> 55
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-PCR fwd

<400> 39
acactctttc cctacacgac gctcttccga tctcattgcc agccattcta gcagg          55


<210> 40
<211> 56
<212> DNA
<213> Artificial Sequence


<220> 
<223> Alternative SARS-CoV-2-PCR rev

<400> 40
tgactggagt tcagacgtgt gctcttccga tctgttgaat ttcttgaact gttgcg         56


