TABLE 5.
Analytical specificity of the qPCR assay for L. plantarum ATCC 202195
| Reaction no. | Contents of reaction | Reaction contained L. plantarum ATCC 202195 | Preparation description | Number of reactions run | Number of reactions that amplified | MeanCq c | Cq range across all reactions |
|---|---|---|---|---|---|---|---|
| 1 | L. crispatus VPI 3199 ATCC 33820DQ (2.5 ng/μL) | No | Single strain | 4 | 0 | d - | - |
| 2 | L. crispatus VPI 3199 ATCC 33820DQ (0.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 3 | L. gasseri DSM 20243 ATCC 33323DQ (2.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 4 | L. gasseri DSM 20243 ATCC 33323DQ (0.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 5 | L. acidophillus ATCC 4357D-5 (2.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 6 | L. acidophillus ATCC 4357D-5 (0.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 7 | L. casei ATCC 334D-5 (2.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 8 | L. casei ATCC 334D-5 (0.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 9 | L. inners AB107 ATCC 55195DQ (2.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 10 | L. inners AB107 ATCC 55195DQ (0.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 11 | L. reuteri F275 ATCC 23272D-5 (2.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 12 | L. reuteri F275 ATCC 23272D-5 (0.5 ng/μL) | No | Single strain | 4 | 0 | - | - |
| 13 | L. plantarum ATCC 14917 (50 ng/μL) | No | Single strain | 3 | 0 | - | - |
| 14 | L. plantarum ATCC 14917 (5 ng/μL) | No | Single strain | 3 | 0 | - | - |
| 15 | L. plantarum ATCC 14917 (2.5 ng/μL) | No | Single strain | 2 | 0 | - | - |
| 16 | L. plantarum ATCC 14917 (0.5 ng/μL) | No | Single strain | 5 | 0 | - | - |
| 17 | L. plantarum ATCC 202195 (50 ng/μL) | Yes | Single strain | 11 | 11 | 17.98 | 17.16, 19.72 |
| 18 | L. plantarum ATCC 202195 (5 ng/μL) | Yes | Single strain | 11 | 11 | 21.66 | 20.44, 22.89 |
| 19 | L. plantarum ATCC 202195 (0.5 ng/μL) | Yes | Single strain | 11 | 10 | 24.83 | 24.07, 26.36 |
| 20 | L. plantarum ATCC 202195 (0.05 ng/μL) | Yes | Single strain | 11 | 11 | 28.25 | 27.32, 30.07 |
| 21 | L. plantarum ATCC 202195 (0.005 ng/μL) | Yes | Single strain | 11 | 11 | 31.7 | 30.93, 33.75 |
| 22 | L. plantarum ATCC 202195 (0.0005 ng/μL) | Yes | Single strain | 11 | 11 | 35.16 | 34.30, 37.05 |
| 23 | L. plantarum ATCC 202195 plus L. crispatus VPI 3199 ATCC 33820DQ (1:1 mixture, 2.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 23.41 | 22.67, 23.86 |
| 24 | L. plantarum ATCC 202195 plus L. crispatus VPI 3199 ATCC 33820DQ (1:1 mixture, 0.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 25.96 | 25.63, 26.17 |
| 25 | L. plantarum ATCC 202195 plus L. gasseri DSM 20243 ATCC 33323DQ (1:1 mixture, 2.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 23.18 | 22.59, 23.72 |
| 26 | L. plantarum ATCC 202195 plus L. gasseri DSM 20243 ATCC 33323DQ (1:1 mixture, 0.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 25.46 | 25.23, 25.75 |
| 27 | L. plantarum ATCC 202195 plus L. acidophillus (1:1 mixture, 2.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 23.1 | 22.73, 23.29 |
| 28 | L. plantarum ATCC 202195 plus L. acidophillus (1:1 mixture, 0.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 25.76 | 25.45, 26.24 |
| 29 | L. plantarum ATCC 202195 plus L. casei (1:1 mixture, 2.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 23.53 | 23.39, 23.91 |
| 30 | L. plantarum ATCC 202195 plus L. casei (1:1 mixture, 0.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 25.49 | 25.19, 25.94 |
| 31 | L. plantarum ATCC 202195 plus L. inners AB107 ATCC 55195DQ (1:1 mixture, 2.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 23.21 | 22.44, 23.80 |
| 32 | L. plantarum ATCC 202195 plus L. inners AB107 ATCC 55195DQ (1:1 mixture, 0.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 25.5 | 25.22, 26.05 |
| 33 | L. plantarum ATCC 202195 plus L. reuteri (1:1 mixture, 2.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 23.52 | 23.27, 24.10 |
| 34 | L. plantarum ATCC 202195 plus L. reuteri (1:1 mixture, 0.5 ng/μL each) | Yes | Multi-strain | 4 | 4 | 25.87 | 25.57, 26.03 |
| 35 | L. plantarum ATCC 202195 plus L. plantarum ATCC 14917 (1:1 mixture, 50 ng/μL each) | Yes | Multi-strain | 3 | 3 | 18.46 | 17.21, 19.83 |
| 36 | L. plantarum ATCC 202195 plus L. plantarum ATCC 14917 (1:1 mixture, 5 ng/μL each) | Yes | Multi-strain | 3 | 3 | 21.48 | 21.35, 21.56 |
| 37 | L. plantarum ATCC 202195 plus L. plantarum ATCC 14917 (1:1 mixture, 2.5 ng/μL each) | Yes | Multi-strain | 2 | 2 | 23.08 | 22.95, 23.22 |
| 38 | L. plantarum ATCC 202195 plus L. plantarum ATCC 14917 (1:1 mixture, 0.5 ng/μL each) | Yes | Multi-strain | 5 | 5 | 25.51 | 24.60, 26.27 |
| 39 | Infant stool samples collected in an observational study conducted in Dhaka, Bangladesh | No | Stool samples | 90 a | 0 | - | - |
| 40 | Infant stool samples collected in an observational study conducted in Dhaka, Bangladesh plus L. plantarum ATCC 202195 cells (2.35 × 107) | Yes | Stool samples | 10 b | 10 | 20.62 | 20.02, 21.21 |
| 41 | Infant stool samples collected in an observational study conducted in Dhaka, Bangladesh plus L. plantarum ATCC 202195 cells (2.35 × 106) | Yes | Stool samples | 10 b | 10 | 23.09 | 21.48, 24.06 |
| 42 | Infant stool samples collected in an observational study conducted in Dhaka, Bangladesh plus L. plantarum ATCC 202195 cells (2.35 × 105) | Yes | Stool samples | 10 b | 10 | 27.34 | 26.25, 28.13 |
a
DNA extracted from 80 stool samples was run once, while DNA extracted from the five stool samples used in the L. plantarum ATCC 202195 cell spike-in assays was run in duplicate.
b
DNA extracted from the five stool samples that were mixed with L. plantarum ATCC 202195 cells was run in duplicate.
c
Cq, cycle quantification number.
d
”-” signifies that data are not applicable in a given cell.