Longitudinal HCMV monitoring and the relation between antiviral therapy and development of mutations in UL97 and UL54
| Patient no. . | Specimens (days after PBSCT) . | PCR . | Viral load* . | Therapy (cumulative days) . | IC50(μmol/L)† . | UL97 mutations . | UL54 mutations . | ||
|---|---|---|---|---|---|---|---|---|---|
| GCV . | PFA . | CDV . | |||||||
| 1 | Leukocyte −183 | + | GCV 0, PFA 0 | Negative | |||||
| Serum −67 | + | GCV 29, PFA 0 | Negative | ||||||
| Plasma −18 | − | GCV 68, PFA 0 | |||||||
| Plasma −12 | + | GCV 68, PFA 0 | Negative | ||||||
| Leukocyte +3 | + | GCV 68, PFA 13 | Negative | ||||||
| Plasma +17 | + | ++ | GCV 68, PFA 27 | Negative | |||||
| Serum +26 | + | ++ | GCV 75, PFA 30 | Negative | |||||
| Leukocyte +32 | + | ++ | GCV 81, PFA 30 | 1.4 | Negative | ||||
| Serum +44 | + | +++ | GCV 93, PFA 30 | C603W | |||||
| Urine +54 | + | +++ | GCV 103, PFA 30 | 13 | 100 | C603W | |||
| Urine +149 | + | GCV 103, PFA 125 | 9.8 | 77 | C603W; A594V | ||||
| 2 | Serum −14 | − | |||||||
| Throat −6 | + | Negative | |||||||
| Leukocyte −1 | + | + | GCV 0, PFA 0 | Negative | |||||
| Leukocyte +6 | + | + | GCV 0, PFA 6 | Negative | |||||
| Leukocyte +42 | + | GCV 24, PFA 18 | Negative | ||||||
| Leukocyte +71 | + | GCV 53, PFA 18 | A591V | ||||||
| Leukocyte +100 | + | ++ | GCV 81, PFA 19 | A591V | Wild type | ||||
| Leukocyte +167 | + | + | GCV 141, PFA 56 | A591V; C603W; M460I | D515E; L516M; I521T‡ | ||||
| Leukocyte +195 | + | ++++ | GCV 156, PFA 56, CDV 14 | A591V | |||||
| Urine +195 | + | ++++ | GCV 156, PFA 56, CDV 14 | 29 | 388 | 7 | A591V | D515E; L516M; I521T; L802M | |
| Leukocyte +251 | + | ++ | GCV 174, PFA 56, CDV 35 | A591V | |||||
| Urine +251 | + | ++ | GCV 174, PFA 56, CDV 35 | 29 | 551 | 7 | A591V; | D515E; L516M; I521T; L802M | |
| Leukocyte +258 | + | GCV 174, PFA 62, CDV 35 | A591V | ||||||
| CSF +263 | + | GCV 179, PFA 67, CDV 35 | P521L‡ | D515E; L516M; I521T | |||||
| A885T; P887S; N898D | |||||||||
| 3 | Plasma −3 | − | GCV 0 | ||||||
| Serum +30 | − | GCV 0 | |||||||
| Leukocyte +65 | + | GCV 0 | Negative | ||||||
| Leukocyte +71 | + | GCV 6 | Negative | ||||||
| Plasma +86 | + | ++ | GCV 21 | Negative | |||||
| Leukocyte +95 | + | GCV 30 | L595S | ||||||
| Plasma +95 | − | GCV 30 | |||||||
| Serum +106 | + | GCV 41 | L595S | ||||||
| Leukocyte +129 | + | GCV 48, CDV 14 | L595S | ||||||
| Leukocyte +169 | + | GCV 48, CDV 35 | Negative | ||||||
| Plasma +224 | − | GCV 48, CDV 35 | |||||||
| Throat +224 | + | GCV 48, CDV 35 | 8 | 240 | 0.3 | L595S | |||
| Patient no. . | Specimens (days after PBSCT) . | PCR . | Viral load* . | Therapy (cumulative days) . | IC50(μmol/L)† . | UL97 mutations . | UL54 mutations . | ||
|---|---|---|---|---|---|---|---|---|---|
| GCV . | PFA . | CDV . | |||||||
| 1 | Leukocyte −183 | + | GCV 0, PFA 0 | Negative | |||||
| Serum −67 | + | GCV 29, PFA 0 | Negative | ||||||
| Plasma −18 | − | GCV 68, PFA 0 | |||||||
| Plasma −12 | + | GCV 68, PFA 0 | Negative | ||||||
| Leukocyte +3 | + | GCV 68, PFA 13 | Negative | ||||||
| Plasma +17 | + | ++ | GCV 68, PFA 27 | Negative | |||||
| Serum +26 | + | ++ | GCV 75, PFA 30 | Negative | |||||
| Leukocyte +32 | + | ++ | GCV 81, PFA 30 | 1.4 | Negative | ||||
| Serum +44 | + | +++ | GCV 93, PFA 30 | C603W | |||||
| Urine +54 | + | +++ | GCV 103, PFA 30 | 13 | 100 | C603W | |||
| Urine +149 | + | GCV 103, PFA 125 | 9.8 | 77 | C603W; A594V | ||||
| 2 | Serum −14 | − | |||||||
| Throat −6 | + | Negative | |||||||
| Leukocyte −1 | + | + | GCV 0, PFA 0 | Negative | |||||
| Leukocyte +6 | + | + | GCV 0, PFA 6 | Negative | |||||
| Leukocyte +42 | + | GCV 24, PFA 18 | Negative | ||||||
| Leukocyte +71 | + | GCV 53, PFA 18 | A591V | ||||||
| Leukocyte +100 | + | ++ | GCV 81, PFA 19 | A591V | Wild type | ||||
| Leukocyte +167 | + | + | GCV 141, PFA 56 | A591V; C603W; M460I | D515E; L516M; I521T‡ | ||||
| Leukocyte +195 | + | ++++ | GCV 156, PFA 56, CDV 14 | A591V | |||||
| Urine +195 | + | ++++ | GCV 156, PFA 56, CDV 14 | 29 | 388 | 7 | A591V | D515E; L516M; I521T; L802M | |
| Leukocyte +251 | + | ++ | GCV 174, PFA 56, CDV 35 | A591V | |||||
| Urine +251 | + | ++ | GCV 174, PFA 56, CDV 35 | 29 | 551 | 7 | A591V; | D515E; L516M; I521T; L802M | |
| Leukocyte +258 | + | GCV 174, PFA 62, CDV 35 | A591V | ||||||
| CSF +263 | + | GCV 179, PFA 67, CDV 35 | P521L‡ | D515E; L516M; I521T | |||||
| A885T; P887S; N898D | |||||||||
| 3 | Plasma −3 | − | GCV 0 | ||||||
| Serum +30 | − | GCV 0 | |||||||
| Leukocyte +65 | + | GCV 0 | Negative | ||||||
| Leukocyte +71 | + | GCV 6 | Negative | ||||||
| Plasma +86 | + | ++ | GCV 21 | Negative | |||||
| Leukocyte +95 | + | GCV 30 | L595S | ||||||
| Plasma +95 | − | GCV 30 | |||||||
| Serum +106 | + | GCV 41 | L595S | ||||||
| Leukocyte +129 | + | GCV 48, CDV 14 | L595S | ||||||
| Leukocyte +169 | + | GCV 48, CDV 35 | Negative | ||||||
| Plasma +224 | − | GCV 48, CDV 35 | |||||||
| Throat +224 | + | GCV 48, CDV 35 | 8 | 240 | 0.3 | L595S | |||
HCMV indicates human cytomegalovirus; PBSCT, peripheral blood stem cell transplantation; PCR, polymerase chain reaction; IC50, 50% inhibitory concentration; GCV, ganciclovir; PFA, foscarnet; CDV, cidofovir; CSF, cerebrospinal fluid.
Limiting-dilution nested PCR from native plasma/serum; + indicates undiluted (corresponding to 103 genome equivalents/mL), ++ indicates dilution of 10; +++ indicates dilution of 100.
Phenotypic resistance testing of viral isolates: cutoff values for resistance: GCV, IC50 > 6 μmol/L; CDV, IC50 > 4 μmol/L; PFA, IC50 > 400 μmol/L.
Mutated codon is located adjacent to known codons involved in drug resistance.