Volume 14, Number 3—March 2008
Research
Discovering and Differentiating New and Emerging Clonal Populations of Chlamydia trachomatis with a Novel Shotgun Cell Culture Harvest Assay
Naraporn Somboonna*†, Sally Mead*, Jessica Liu†, and Deborah Dean*†‡
Table 1
Results of the modified plaque assay for Chlamydia reference strains and shotgun cell culture harvest technique for clinical strains representing acute and persistent infections*
| Strain | Days p.i. to plaque formation or harvest | No. plaques or shotgun harvested areas | ompA genotype (no.) | Location of nucleotide substitutions in ompA (amino acid substitution location) | 16S rRNA (no.) |
|---|---|---|---|---|---|
| Reference D/UW-3 and E/Bour 1:1† | 9 | 25 | D (13) E (9) D/E (3) | – – – | Chlamydia trachomatis D (13) C. trachomatis E (9) C. trachomatis D/E (3) |
| Reference D/UW-3 and E/Bour 3:‡ | 9 | 9 | D (9) E (0) | – – | C. trachomatis D (9) C. trachomatis E (0) |
| D/UW-3/E/Bour mixed infection§ | 9 | 11 | D (7) E (4) | C. trachomatis D (7) C. trachomatis E (4) | |
| Acute clinical Ja | 8 | 11 | Ja (11) | – | C. trachomatis Ja (11) |
| Acute clinical K | 10 | 11 | K (11) | – | C. trachomatis K (11) |
| Persistent H | 7 | 5 | H (5) | – | C. trachomatis H (5) |
| Persistent G | 14 | 7 | G (7) | – | C. trachomatis G (7) |
| Persistent F | 10 | 5 | F (5) | – | C. trachomatis F (5) |
| Persistent clinical F and G strains 1:1† | 10 | 13 | F (1) G (12) | – – | C. trachomatis F (1) C. trachomatis G (12) |
| A/SA-1 | 10 10¶ | 18 21 | A (14) Chlamydophila abortus (4) A/C. abortus (21) | – – | C. trachomatis A (14) C. abortus (4) C. trachomatis A/C. abortus (21) |
| B/TW-5 | 9 | 15 | B (15) | – | C. trachomatis B (15) |
| Ba/Apache-2 | 8 | 14 | Ba (9) Ba1 (1) Ba2 (1) Ba3 (1) | – C662T (P221L) C662T (P221L) G673A (E225K) C662T (P221L) A717C (K239N) | C. trachomatis Ba (14) |
| C/TW-3 | 12 | 13 | C (13) | – | C. trachomatis C (13) |
| D/UW-3 | 9 | 11 | D (11) | – | C. trachomatis D (11) |
| Da/TW-448 | 10 | 12 | Da (12) | – | C. trachomatis Da (12) |
| E/Bour | 9 | 9 | E (9) | – | C. trachomatis E (9) |
| F/IC-Cal3 | 9 | 13 | F (10) F-III (3) | – G269A (G90E)# | C. trachomatis F (13) |
| G/UW57/Cx | 7 | 10 | G (10) | – | C. trachomatis G (10) |
| H/UW-4 | 12 | 12 | H (10) | – | C. trachomatis H(10) |
| I/UW-12 | 11 | 14 | I (14) | – | C. trachomatis I (14) |
| Ia/IU-4168 | 12 | 11 | Ia (12) | – | C. trachomatis Ia (12) |
| J/UW-36 | 11 | 15 | J (15) | – | C. trachomatis J (15) |
| Ja/UW-92 | 12 | 12 | Ja (12) | – | C. trachomatis Ja (12) |
| K/UW-31 | 11 | 13 | K (13) | – | C. trachomatis K (13) |
| L1/440 | 9 | 11 | L1 (11) | – | C. trachomatis L1 (11) |
| L2/434 | 9 | 11 | L2 (8) L2’ (3) | – C471G, G496A (A166T)¶ | C. trachomatis L2 (10) C. trachomatis L2a (1) |
| L2a/TW-396 | 9 | 13 | L2a (13) | – | C. trachomatis L2 (13) |
| L3/404 | 9 | 14 | L3 (14) | – | C. trachomatis L3 (14) |
*p.i.,.postinoculation; ompA, outer membrane protein A; –, no evidence for nucleotide substitution.
†Represents an equal mixture of inclusion-forming units (IFUs) of each strain.
‡Represents a 3:1 mixture of IFUs of reference strains D/UW-3 and E/Bour.
§Represents the plaques from the 1:1 mixture of D/UW-3 and E/Bour when both reference strains were found in 3 plaques.
¶Represents the plaques that were harvested from the wells with 10–8 dilution.
#Boldface denotes nonconservative amino acid substitution (17).
