Background
Methods
Materials
Generation of recombinant gp160 antigens
Preparation of labeled antigens
In-house TRF double antigen sandwich assays
Data analysis
Results and discussion
Recombinant gp160 antigens
Design of double antigen sandwich TRF assays
Performance evaluation of the r-Δgp160-based one- and two-step double antigen sandwich TRF assays
Member ID# | Days since 1stbleed | Abbott | Gen. Sys. | Organon Teknika HIVa
| TRF assaysb
| |||
---|---|---|---|---|---|---|---|---|
HIV1a
| HIV1/2a
| HIV1a
| HIV1/2a
| r-Δgp160 two-step | r-Δgp160 one-step | |||
01 | 0 | 0.2 | 0.1 | 0.2 | 0.1 | 0.3 | 0.9 (−) | 0.5 (−) |
02 | 2 | 0.2 | 0.1 | 0.1 | 0.1 | 0.3 | 0.8 (−) | 0.5 (−) |
03 | 7 | 0.2 | 0.1 | 0.2 | 0.1 | 0.4 | 0.8 (−) | 0.4 (−) |
04 | 9 | 0.2 | 0.1 | 0.2 | 0.1 | 0.3 | 0.8 (−) | 0.5 (−) |
05 | 15 | 0.2 | 0.1 | 0.2 | 0.1 | 0.3 | 0.7 (−) | 0.5 (−) |
06 | 28 | 0.9 | 6 | 0.3 | 0.4 | 0.6 | 17.2 (+) | 5.6 (+) |
07 | 33 | 3.9 | >18.7 | 0.8 | 1.1 | 2.3 | 15.9 (+) | 4.3 (+) |
08 | 35 | 5.7 | >18.7 | 1.3 | 1.9 | 3.1 | 11.9 (+) | 3.3 (+) |
09 | 42 | 10.5 | >18.7 | 2.9 | 4 | 4.6 | 13.8 (+) | 2.9 (+) |
Member ID# | Origin | Gtpa
| Abbott | Gen. Sys. | OT HIV1b
| TRF assaysc
| |||
---|---|---|---|---|---|---|---|---|---|
HIV1b
| HIV1/2b
| HIV1b
| HIV1/2b
| r-Δgp160 two-step | r-Δgp160 one-step | ||||
01 | Spain | O | 1.1 | 1.8 | 0.8 | 5.6 | 1.3 | 1.3 (+) | 0.6 (−) |
02 | Ghana | A | >11.5 | >16.1 | 6.9 | 8.7 | 7.0 | 94.0 (+) | 9.2 (+) |
03 | Ghana | G | >11.5 | >16.1 | 7.1 | 8.8 | 7.2 | 164.8 (+) | 41.6 (+) |
04 | Ghana | G | >11.5 | >16.1 | 7.1 | 8.8 | 6.5 | 138.0 (+) | 4.1 (+) |
05 | Ghana | A | >11.5 | >16.1 | 7.1 | 8.7 | 7.0 | 253.6 (+) | 82.2 (+) |
06 | Ghana | G | >11.5 | >16.1 | 6.9 | 8.8 | 7.1 | 91.4 (+) | 15.1 (+) |
08 | Ivory Coast | G | >11.5 | >16.1 | 6.9 | 8.7 | 6.7 | 73.0 (+) | 4.4 (+) |
09 | Ivory Coast | A | >11.5 | >16.1 | 6.9 | 8.6 | 6.5 | 107.8 (+) | 3.9 (+) |
10 | Ivory Coast | Neg | 0.4 | 0.2 | 0.1 | 0.4 | 0.4 | 0.7 (−) | 0.4 (−) |
11 | Mozambique | HIV-2 | 1.2 | 14.6 | 0.6 | 9.7 | 3.0 | 1.0 (+) | 0.7 (−) |
12 | Mozambique | C | >11.5 | >16.1 | 7.1 | 8.9 | 6.9 | 187.1 (+) | 4.1 (+) |
14 | Uganda | D | >11.5 | >16.1 | 4.5 | 8.5 | 6.2 | 53.3 (+) | 13.8 (+) |
15 | Uganda | D | >11.5 | >16.1 | 6.3 | 8.1 | 7.2 | 132.9 (+) | 16.8 (+) |
16 | Uganda | D | >11.5 | >16.1 | 7.0 | 8.8 | 6.9 | 154.5 (+) | 7.3 (+) |
17 | Uganda | D | >11.5 | >16.1 | 6.8 | 9.8 | 7.0 | 131.2 (+) | 5.1 (+) |
19 | Zimbabwe | C | >11.5 | >16.1 | 6.0 | 9.9 | 7.0 | 41.4 (+) | 10.3 (+) |
21 | China | B | >11.5 | >16.1 | 6.7 | 8.8 | 7.0 | 136.7 (+) | 26.8 (+) |
22 | Thailand | E | >11.5 | >16.1 | 7.3 | 9.8 | 7.0 | 100.8 (+) | 7.0 (+) |
24 | Thailand | E | >11.5 | >16.1 | 7.4 | 9.8 | 6.9 | 88.4 (+) | 23.2 (+) |
25 | India | HIV-2 | 0.4 | 15.4 | 3.8 | 10 | 2.1 | 4.9 (+) | 1.5 (+) |
26 | USA | D | >11.5 | >16.1 | 7.4 | 9.8 | 7.1 | 85.8 (+) | 48.5 (+) |
27 | USA | B/D | >11.5 | >16.1 | 7.0 | 9.8 | 7.2 | 135.7 (+) | 12.7 (+) |
28 | Argentina | F | >11.5 | >16.1 | 7.0 | 8.9 | 6.8 | 72.5 (+) | 3.0 (+) |
29 | Argentina | B | >11.5 | >16.1 | 6.9 | 8.5 | 6.6 | 92.9 (+) | 2.2 (+) |
30 | Argentina | Neg | 0.3 | 0.2 | 0.2 | 0.2 | 0.4 | 0.9 (−) | 0.4 (−) |
Member ID# | Ortho EIA HCVa
| Abbott EIA HBsAga
| OT Anti-HBca
| HTLV | HIV-1 | TRF assaysb
| |||
---|---|---|---|---|---|---|---|---|---|
Abbott EIAa
| GS Blotsa
| Abbott EIAa
| Dupont blotsa
| r-Δgp160 two-step | r-Δgp160 one-step | ||||
1 | + | + | + | + | P | 0.2 | N/A | 0.9 (−) | 0.6 (−) |
2 | - | + | + | - | N/A | 13.5 | P | 96.5 (+) | 6.5 (+) |
3 | + | + | + | + | P | 0.1 | N/A | 0.5 (−) | 0.7 (−) |
4 | - | + | + | + | P | 0.1 | N/A | ND | ND |
5 | + | - | + | + | P | 13.5 | P | 122.6 (+) | 8.5 (+) |
6 | - | - | - | - | N/A | 0.1 | N/A | ND | ND |
7 | + | - | - | + | P | 13.5 | P | 146.1 (+) | 9.0 (+) |
8 | + | + | + | - | N/A | 13.5 | P | 208.4 (+) | 5.2 (+) |
9 | + | + | - | + | P | 13.5 | P | 107.7 (+) | 8.5 (+) |
10 | - | + | + | - | N/A | 13.5 | P | 84.9 (+) | 13.3 (+) |
11 | + | + | + | + | P | 0.1 | N/A | 0.7 (−) | 0.6 (−) |
12 | + | + | + | - | N/A | 13.5 | P | 142.2 (+) | 1.1 (+) |
13 | - | + | + | - | N/A | 13.5 | P | 47.8 (+) | 11.2 (+) |
14 | - | + | + | + | P | 0.32 | N/A | 0.8 (−) | 0.7 (−) |
15 | + | + | + | + | IND | 0.2 | N/A | 0.8 (−) | 0.7 (−) |
16 | + | - | + | + | P | 13.5 | P | 182.5 (+) | 4.5 (+) |
17 | + | + | + | - | N/A | 13.5 | P | 215.4 (+) | 9.1 (+) |
18 | + | + | - | + | P | 13.5 | P | 48.3 (+) | 3.6 (+) |
19 | - | + | + | - | N/A | 13.5 | P | 88.2 (+) | 9.7 (+) |
20 | - | + | + | - | N/A | 1.1 | P | 3.3 (+) | 2.5 (+) |
21 | - | + | + | - | N/A | 13.5 | P | 128.6 (+) | 3.0 (+) |
22 | + | + | + | - | N/A | 13.5 | P | 140.6 (+) | 16.9 (+) |
23 | + | + | + | + | P | 0.2 | N/A | 0.8 (−) | 0.8 (−) |
24 | - | - | - | - | N/A | 0.1 | N/A | 0.8 (−) | 0.8 (−) |
25 | - | + | + | - | N/A | 13.5 | P | 127.1 (+) | 6.0 (+) |
Member ID # | Abbott HIV 1/2a
| Gen. Sys. HIV 1/2a
| Org. Tek. HIV 1a
| Bio-Rad HIV-1 Western Blot Resultb
| Roche PCR HIV-1 RNA test copies/mlc
| TRF assaysd
| ||
---|---|---|---|---|---|---|---|---|
r-Δgp160 two-step | r-Δgp160 one-step | r-HIV-1env two-step | ||||||
1 | 5.5 | >8.9 | 1.4 | + | 3x104
| 1.9 (+) | 1.8 (+) | 2.0 (+) |
2 | 0.2 | 0.3 | 0.3 | - | BLD | 0.8 (−) | 0.6 (−) | 0.7 (−) |
3e
| NA | NA | 1.8 | IND | 5x101* | 3.4 (+) | 3.0 (+) | 0.8 (−) |
4e
| 11.9 | >8.9 | 6.3 | + | 9x104
| 7.0 (+) | 5.2 (+) | 3.4 (+) |
5 | 3.4 | >8.9 | 3.2 | + | 5x101* | 2.2 (+) | 1.2 (+) | 3.6 (+) |
6 | NA | NA | 1.2 | IND | 6x103
| 1.8 (+) | 1.1 (+) | 5.9 (+) |
7f
| 5.3 | >8.9 | 1.3 | + | 3x103
| 2.7 (+) | 1.6 (+) | 3.9 (+) |
8f
| 8.0 | >8.9 | 2.8 | + | 7x103
| 3.8 (+) | 2.8 (+) | 4.8 (+) |
9 | 8.5 | >8.9 | 0.7 | + | 2x103
| 1.9 (+) | 1.1 (+) | 7.2 (+) |
10 | 8.6 | >8.9 | 0.6 | IND | 3x105
| 1.7 (+) | 1.2 (+) | 13.2 (+) |
11 | NA | NA | 3.8 | + | 8x103
| 3.2 (+) | 2.0 (+) | 5.4 (+) |
12 | 1.7 | 2.0 | 0.3 | - | 5x105
| 1.1 (+) | 0.6 (−) | 0.9 (−) |
13 | NA | NA | 0.9 | IND | 2x105
| 4.9 (+) | 2.7 (+) | 18.2 (+) |
14g
| NA | NA | 0.4 | - | 3x105
| 1.4 (+) | 1.0 (+) | 1.1 (+) |
15g
| 14.2 | >8.9 | 1.8 | IND | >7x105
| 5.9 (+) | 3.0 (+) | 8.0 (+) |