Sample ID Culture (24-48 hr) RV-PCR (9 hr)
Average CFU/Sample Source of Culture Result* PCR of Culture (Ct)** Culture Result (Pos/Neg) Average ΔCt Std Dev ΔCt RV-PCR Results (Pos/Neg) Notes
1185 0 EC-PE 25.2 Pos 7.1 1.8 Pos See Footnote (1)
1198 0 N/A N/A Neg 0.0 0.0 Neg  
1199 0 N/A N/A Neg 0.0 0.0 Neg  
1243 3.0E1 Plates 15.7 Pos 6.5 0.9 Pos  
1246 3.0E1 Plates 13.7 Pos 6.5 0.9 Pos  
1255 0 EC 25.6 Pos 5.6 0.3 Neg RV-PCR Ave. ΔCt from 1:20 dilution
1257 3.0E1 Plates 14.6 Pos 6.1 1.3 Pos  
1259 0 EC 21.9 Pos 0.0 0.0 Neg Heat Lysis Ave. ΔCt = 3.7
1261 0 EC-PE 25.8 Pos 7.8 1.2 Pos See Footnote (1)
1290 0 EC-PE 23.8 Pos 8.6 0.6 Pos See Footnote (1)
1291 0 EC 25.6 Pos 10.9 0.1 Pos See Footnote (2)
1292 0 EC 25.5 Pos 12.9 0.2 Pos See Footnote (2)
1298 1.3E3 Plates, FF 12.9 Pos 8.3 0.7 Pos  
1300 3.0E1 Plates 13.6 Pos 6.7 0.8 Pos  
1301 0 EC 25.4 Pos 4.2 2.0 Pos Ave. ΔCt from 1:20 dilution; Heat Lysis Ave. ΔCt = 8.4
1302 0 N/A N/A Neg 0.0 0.0 Neg  
1303 0 EC 35.0 Pos 9.6 2.2 Pos  
1304 0 EC 23.8 Pos 14.8 0.1 Pos See Footnote (2)
1305 5.3E1 Plates, FF 13.3 Pos 20.2 1.8 Pos  
1306 1.8E1 FF, RS 14.5 Pos 16.0 1.7 Pos  
1475 (TB) 0 N/A N/A Neg 0.0 0.0 Neg  
1476 (TB) 0 N/A N/A Neg 0.0 0.0 Neg  
NC 0 N/A N/A Neg 0.0 0.0 Neg  
PC 2.9E5 Plates 14.0 Pos 23.6 0.1 Pos  
1307 0 EC 35.0 Pos 6.7 0.4 Pos  
1308 8.9E1 Plates, FF 13.6 Pos 7.9 0.6 Pos  
1309 0 N/A N/A Neg 0.0 0.0 Neg  
1310 0 EC 16.2 Pos 8.7 0.3 Pos  
1311 0 N/A Undetermined Neg 6.4 0.3 Pos RV-PCR Ave. ΔCt from 2 replicates (1:20 dilution)
1312 0 EC 26.0 Pos 7.7 0.3 Pos  
1374 0 N/A N/A Neg 0.0 0.0 Neg  
1375 0 N/A N/A Neg 0.0 0.0 Neg  
1377 0 N/A N/A Neg 0.0 0.0 Neg  
1379 0 EC-PE 24.8 Pos 12.3 0.5 Pos See Footnote (3)
1382 0 EC-PE 27.0 Pos 12.6 0.1 Pos See Footnote (3)
1383 0 N/A N/A Neg 0.0 0.0 Neg  
NC 0 N/A N/A Neg 0.0 0.0 Neg  
PC 4.7E5 Plates 14.0 Pos 24.9 0.1 Pos  
1163 0 EC Undetermined Neg 0.0 0.0 Neg  
1164 0 RS 15.0 Pos 5.9 0.6 Pos See Footnote (4): Heat Lysis Ave. ΔCt = 8.3
1165 5.9E1 Plates 14.7 Pos 8.8 0.2 Pos  
1168 0 RS 14.9 Pos 6.1 0.4 Pos  
1171 0 N/A N/A Neg 0.0 0.0 Neg  
1177 0 RS 15.2 Pos 8.6 0.7 Pos  
1179 0 RS 14.7 Pos 4.9 1.0 Pos See Footnote (4): Heat Lysis Ave. ΔCt 8.7
1180 0 FF, RS 34.6 Pos 9.3 0.3 Pos  
1181 0 EC Undetermined Neg 0.0 0.0 Neg  
1182 3.0E1 Plates, FF 14.4 Pos 7.0 0.8 Pos  
1186 0 EC 19.9 Pos 7.5 0.8 Pos  
1210 3.6E1 FF, EC 14.1 Pos 6.9 2.0 Pos  
1464 (TB) 0 N/A N/A Neg 0.0 0.0 Neg  
1465 (TB) 0 N/A N/A Neg 0.0 0.0 Neg  
1466 (TB) 0 N/A N/A Neg 0.0 0.0 Neg  
NC 0 N/A N/A Neg 0.0 0.0 Neg  
PC 3.4E5 Plates 13.5 Pos 20.6 0.1 Pos  
*Positive (pos) or negative (neg) culture determination based upon the serial dilution (plates), filter funnel (FF) plate, and/or enrichment culture (EC), re-streak (RS) from enrichment culture or Promega-extracted concentrated EC (EC-PE). Positive culture results obtained if > 0 Bg colonies were present on serial dilution or filter membrane plates, and/or Bg-positive PCR results were obtained (Ct < 35) from colonies and/or the EC or RS. N/A = not applicable; No CFU for analysis and EC not turbid and/or no growth from RS. **Cycle threshold (Ct) values obtained from real-time PCR analysis of selected colonies and/or enrichment culture (EC). Positive RV-PCR result based upon average ΔCt≥ 6 and T9 Ct ≤ 39. Footnote (1): EC concentrated 10- to 20-fold and DNA prepared using Promega extraction prior to PCR analysis to confirm culture result (EC-PE). Footnote (2): PCR repeated to confirm RV-PCR result; 1:20 dilution result shown. Footnote (3): RV-PCR results based on repeated PCR analysis with 1:20 dilution (1:10 dilution showed PCR inhibition); EC concentrated 10- to 20-fold and DNA prepared using Promega extraction prior to PCR to confirm culture result (EC-PE). Footnote (4): Low spore level, post-decontamination sample; sample positive by heat lysis RV-PCR. Abbreviations: PC, positive control; NC, negative control; TB, true blank.
Table 4: RV-PCR and culture results for VHP post-decontamination samples.