<p> 1Title: Validation of a Duplex Real-Time PCR for the Detection of Salmonella spp. in </p><p>2different Food Products</p><p>3Authors: A. Anderson1, K. Pietsch1, R. Zucker2, A. Mayr2, E. Müller-Hohe1, U. </p><p>4Messelhäusser2, A. Sing2, U. Busch2, and I. Huber2</p><p>51Chemisches und Veterinäruntersuchungsamt (CVUA), Bissierstr. 5, 79114 Freiburg, </p><p>6Germany</p><p>72Bavarian Health and Food Safety Authority (LGL), Oberschleißheim, Germany</p><p>8Running title: Real-time PCR for Salmonella spp. in food</p><p>9Correspondence to: Dr. Ingrid Huber, Bavarian Health and Food Safety Authority, </p><p>10Veterinärstr. 2, D-85764 Oberschleißheim, Germany, Tel: +49-89-31560-158, Fax: </p><p>11+49-89-31560-458</p><p>12</p><p>13</p><p>14</p><p>15</p><p>16</p><p>17</p><p>18</p><p>19</p><p>20</p><p>21</p><p>22</p><p>23</p><p>24</p><p>25</p><p>26</p><p>27Supplementary material 1 1 28Table 1 </p><p>29Salmonella strains used for inclusivity testing of the invA real-time PCR </p><p>Salmonella Sub- Number of PCR Strain ID Serotype speci strains Result es tested</p><p>S. Agona I 2 + bgvv: 169/97; 99-00029 bgvv: 29/97; 99-00929; 99-02762 S. Anatum I 4 + GSA: 9/202 + bgvv: 48;g;251; 99-01556; 98-01776-1; S. Arizonae IIIa 6 98-01776-2; 00-2040; 00-00269 + bgvv: 48;235; bgVV 66;z;65; 96-00845; S. bongori 4 K-1354 bgvv: 91/97; 01-01058 S. Bovismorbificans I 2 + S. Brandenburg I 2 + bgvv: 5/97; 00-00706 S. Broughton I 1 + bgvv: 99-00019 S. Derby I 1 + bgvv: 6/97 + bgvv: 98-00092; 98-02723; 01-01303; S. Diarizonae IIIb 6 01-01515; 01-00631; 01-01749 S. Dublin I 2 + bgvv: 1294/97; GSA9/223 S. Enterica I 1 + GSA: 9/224 S. Enteritidis I 3 + GSA: 9/225; bgVV: 332/97; 00-00004 S. Falkensee I 1 + bgvv: 01-00367 S. Give I 1 + bgvv: 99-01625 S. Goldcoast I 2 + bgvv: 01-01443; 01-02132 S. Hadar I 2 + bgvv: 132/97; 99-00601 S. Heidelberg I 2 + bgvv: 01-00210; 00-00935 S. Houtenae IV 3 + bgvv: 42;236; 98-003903; 98-03722 S. Indica VI 3 + bgvv: 41;b;1,7;lac+; 94-02957; K 934 GSA: 9/240; I + bgvv 480/97; 01-00280; 01-02719; 99- S. Infantis 5 01857 S. Kedougou I 1 + bgvv: 01-00319 S. Kentucky I 1 + bgvv: 01-00936 S. Kodjovi I 1 + bgvv: 98-03226 S. Litchfield I 2 + bgvv: 01-1173; 01-1174 S. Liverpool I 1 + bgvv: 00-00951 2 3 S. Livingstone I 1 + bgvv: 82/97 S. Manhattan I 1 + bgvv: 810/97 S. Montevideo I 1 + bgvv: 106/97 S. Morningside I 1 + bgvv: 98-00220 S. Newport I 2 + bgvv: 99-00095; 107/97 S. Oranienburg I 2 + GSA: 9/179; 9/180 S. Paratyphi I 2 + bgvv: 119/97; 98-02313 S. Panama I 2 + bgvv: 388/97; GSA9/257 S. Pullorum I 1 + bgvv: 341/97 S. Rubislaw I 1 + bgvv: 01-02200 + bgvv : 42;r; 98-01308; 00-01121; 99- S. Salamae II 5 01050; 00-01588; S. Saint-Paul I 1 + bgvv: 00-02151 S. Senftenberg I 4 + bgvv: 01-3212 S. Typhi I 4 + GSA: 9/266; 9/267 ATCC13311; GSA: 9/23; 9/269; 9/65; 9/32 S. Typhimurium I 8 + bgvv: 01-02893; 99-00443; 96-51K61 S. Virchow I 3 + bgvv: 142/97; 01-00826; 00-01570 S. Vittzin I 1 + bgvv: 28;w;enx S. Weltevreden I 2 + bgvv: 99-03668; 01-00926 S. Yoruba I 2 + bgvv: 01-00761; 01-01119 S. Zanzibar 1 + bgvv: 99-04414 30bgvv: strain collection of BfR, Berlin, Germany; 31GSA: strain collection of GeneScan Analytics GmbH, Freiburg, Germany 32 33 34</p><p>3 5 35 36Table 2 37Non-Salmonella species used for exclusivity testing of the invA real-time PCR </p><p>Species Number of PCR Strain ID strains result tested</p><p>Campylobacter coli 1 - DSMZ 4689 Campylobacter jejuni 1 - DSMZ 4688 Campylobacter laridis 1 - GSA: 42/284 Cedecea davisae 1 - DSMZ 4568 Citrobacter amalonaticus 1 - DSMZ 4593 DSMZ 30040; DSMZ 30039; Citrobacter freundii 4 - GSA: 11/97; 11/81 GSA: 10/181; 10/78; 10/171; 10/42; 10/29; 10/30; 10/31; 10/68; 10/69; 10/70; 10/72; Escherichia coli 13 - 10/108; 10/109 Edwardsiella tarda 1 - DSMZ 30052 Enterobacter aerogenes 1 - GSA : 44/286 Enterobacter cloacae 1 - DSMZ 30054 Enterobacter sakazakii 2 GSA : 46/288; 46/289 Enterobacter tarda 1 - GSA: 47/290 Ewingella americana 1 - DSMZ 4580 Hafnia alvei 1 - GSA: 49/292 Klebsiella pneumoniae - GSA: 50/293 ssp. pneumoniae 1</p><p>Klebsiella pneumoniae ssp. ozeanae 1 - GSA: 50/294 Klebsiella oxytoca 1 - GSA: 51/295 Listeria grayi 1 - GSA: 8/12 Listeria innocua 1 - GSA: 4/6 Listeria ivanovii 1 - GSA: 7/11 Listeria monocytogenes 2 - DSMZ 20600; GSA: 3/4 Listeria seeligeri 1 - GSA: 6/10 Listeria welshimeri 1 - GSA: 5/9 Moellerella wisconsensis 1 - DSMZ 5076 Pantoea agglomerans 1 - DSMZ 3493</p><p>4 7 Proteus mirabilis 1 - GSA: 54/298 Proteus vulgaris 1 - GSA: 55/299 Providencia stuartii 1 - DSMZ 4539 Serratia marcescens 1 - DSMZ 30121 Shigella boydii 1 - GSA: 57/301 Shigella dysenteriae 1 - GSA: 58/302 Shigella flexneri 1 - GSA: 59/303 Shigella sonnei 1 - GSA : 60/304 Staphylococcus aureus 1 - GSA : 13/198 Vibrio cholerae 1 - GSA: 61/305 Vibrio damsella 1 - GSA: 62/306 Vibrio mimicus 1 - GSA: 63/307 Vibrio parahaemolyticus 1 - GSA: 64/308 DSMZ 4780; Yersinia enterocolitica 3 - GSA: 65/310; 65/311 Yersinia intermedia 1 - GSA: 66/312 Yersinia - GSA: 67/313 pseudotuberculosis 1 38DSMZ: German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany) 39GSA: strain collection of GeneScan Analytics GmbH, Freiburg, Germany 40 41 42 43 44 45</p><p>46</p><p>5 9</p>