TY - JOUR
T1 - Gene typing of chlamydia trachomatis by polymerase chain reaction and restriction endonuclease digestion
AU - Gaydos, Charlotte A.
AU - Bobo, Linda
AU - Welsh, Laura
AU - Hook, Edward W.
AU - Viscidi, Raphael
AU - Quinn, Thomas C.
PY - 1992
Y1 - 1992
N2 - A portion of the major outer membrane protein (MOMP) gene from 15 Chlamydia trachomatis serovars was amplified by polymerase chain reaction (PCR) and the product was analyzed by restriction fragment length polymorphism (RFLP). A set of primers was used to amplify an 871 base pair gene fragment encompassing the 4 hypervariable regions of MOMP. AluI digestion of the product gave distinctive patterns for the 15 serovars as demonstrated on silver-stained polyacrylamide gels. A triple digest with EcoRI, HinfI, and HpaII allowed improved discrimination between closely related serovars (C, H, I, J, L3). PCR and RFLP were used to type 50 wild-type clinical isolates and results were compared to results of the solid-phase enzyme immunoassay typing method. These isolates represented the most prevalent genital serovars (D, E, F, K, I and J) in the local sexually transmitted diseases clinic population. For specimens containing 1 serovar, the results of the two methods were similar for 42 samples and discordant for 1 sample. In addition, two samples showed evidence of mixed infection with two serovars as identified by both methods. Five additional specimens contained two serovars, as shown by one or both methods. In all five such specimens, the two typing methods agreed on at least one of the two serovars. For both single and multiple serovar specimens, there was concordance between the two typing methods for 16/17 E serovars, 8/9 D serovars, 8/8 F serovars, 7/7 I serovars, 7/7 J serovars, 5/8 K serovars, and 0/2 G serovars. This rapid and economic method can detect mixed infections, may circumvent the need to culture chlamydial organisms for typing, and should be a useful addition to chlamydia typing technology.
AB - A portion of the major outer membrane protein (MOMP) gene from 15 Chlamydia trachomatis serovars was amplified by polymerase chain reaction (PCR) and the product was analyzed by restriction fragment length polymorphism (RFLP). A set of primers was used to amplify an 871 base pair gene fragment encompassing the 4 hypervariable regions of MOMP. AluI digestion of the product gave distinctive patterns for the 15 serovars as demonstrated on silver-stained polyacrylamide gels. A triple digest with EcoRI, HinfI, and HpaII allowed improved discrimination between closely related serovars (C, H, I, J, L3). PCR and RFLP were used to type 50 wild-type clinical isolates and results were compared to results of the solid-phase enzyme immunoassay typing method. These isolates represented the most prevalent genital serovars (D, E, F, K, I and J) in the local sexually transmitted diseases clinic population. For specimens containing 1 serovar, the results of the two methods were similar for 42 samples and discordant for 1 sample. In addition, two samples showed evidence of mixed infection with two serovars as identified by both methods. Five additional specimens contained two serovars, as shown by one or both methods. In all five such specimens, the two typing methods agreed on at least one of the two serovars. For both single and multiple serovar specimens, there was concordance between the two typing methods for 16/17 E serovars, 8/9 D serovars, 8/8 F serovars, 7/7 I serovars, 7/7 J serovars, 5/8 K serovars, and 0/2 G serovars. This rapid and economic method can detect mixed infections, may circumvent the need to culture chlamydial organisms for typing, and should be a useful addition to chlamydia typing technology.
UR - http://www.scopus.com/inward/record.url?scp=0026620351&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026620351&partnerID=8YFLogxK
U2 - 10.1097/00007435-199211000-00001
DO - 10.1097/00007435-199211000-00001
M3 - Article
C2 - 1362824
AN - SCOPUS:0026620351
SN - 0148-5717
VL - 19
SP - 303
EP - 308
JO - Sexually transmitted diseases
JF - Sexually transmitted diseases
IS - 6
ER -