DETECTION AND STRAIN DIFFERENTIATION OF CHLAMYDIA-PSITTACI MEDIATED BY A 2-STEP POLYMERASE CHAIN-REACTION

被引:52
作者
KALTENBOECK, B [1 ]
KOUSOULAS, KG [1 ]
STORZ, J [1 ]
机构
[1] LOUISIANA STATE UNIV,SCH VET MED,DEPT VET MICROBIOL & PARASITOL,BATON ROUGE,LA 70803
关键词
D O I
10.1128/JCM.29.9.1969-1975.1991
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Specific and sensitive amplification of major outer membrane protein (MOMP) gene DNA sequences of Chlamydia psittaci was achieved in a two-step polymerase chain reaction. First, oligonucleotide primers specific for 5' and 3' nontranslated regulatory regions of the MOMP gene were used in a polymerase chain reaction to amplify a DNA fragment of approximately 1,400 bp. A portion of this DNA fragment was amplified in a second reaction using a degenerate oligonucleotide primer specific for a DNA sequence contained within the 1,400-bp DNA fragment and one of the first-step primers. This method detected 10 cognate chlamydial genomes. C. psittaci MOMP genes from two avian strains and from mammalian serovars 1, 7, and 8 were amplified and analyzed by restriction endonuclease digestion. MOMP genes from mammalian serovars 2 through 6 and 9 and from strains of C. trachomatis and C. pneumoniae could not be amplified. Restriction endonuclease analysis with HaeIII indicated a close relationship between C. psittaci strains of avian and mammalian serovar 1 lineage, while those of mammalian serovars 7 and 8 exhibited distinct restriction patterns. DNA sequences corresponding to the mammalian serovar 1-wild type parakeet MOMP genotype of C. psittaci were detected in two of seven milk samples from cases of bovine mastitis.
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页码:1969 / 1975
页数:7
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