Detection of Chlamydia psittaci using DNA probes and the polymerase chain reaction

Rasmussen, S J

doi:10.1016/0378-1097(91)90546-m

Cited by 6 publications

(6 citation statements)

References 9 publications

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“…The sensitivity of detection, estimated at about 2.5 X 106 copies, is low but in relative agreement with data published for other non-isotopic probes specific for different intracellular prokaryotes, like mollicutes (Razin et al 1987), chlamydias (Rasmussen & Timms 1991) or rickettsias (Goff et al 1988). The sensitivity limlts are generally between 0.01 and 1 ng of genomic DNA, which is eqiiivaleiit to about IS3 : a ?G6 s q a n isms.…”

Section: Discussionsupporting

confidence: 67%

Genomic DNA cloning of rikettsia-like organisms (RLO) of Saint-Jacques scallop Pecten maximus: evaluation of pro-karyote diagnosis by hybridization with a non-isotopically labelled probe and by polymerase chain reaction

Kellner-Cousin¹,

Gall²,

Despres³

et al. 1993

Dis. Aquat. Org.

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Genomic DNA fragments of a rickettsia-like organism (RLO) of Saint-Jacques scallop, Pecten maximus, were extracted from purified RLO and cloned. A DNA sequence of 1500 bp taken from one of the sequenced clones was coupled to horseradish peroxidase and the resulting probe was assayed for the diagnosis of RLO by nucleic acid hybridization. The specificity of the probe for RLO DNA was established from its lack of hybridization with scallop DNA. The sensitivity limit of the probe detected by enhanced chemiluminescence was determined to be around 500 ng of total RLO nucleic acids, equivalent to about 2.5 X 106 copies of RLO DNA. A palr of polymerase chain reaction (PCR) primers were synthesized that amplified a 1300 bp segment of the 1500 bp RLO fragment. After 30 cycles of amplification of total RLO nucleic acids, a product of the expected size was easily visible on agarose at all starting template concentrations between 100 ng and 10 pg of RLO total nucleic acids. The lowest level of sensitivity corresponds to about 100 fg of RLO DNA or about 50 genomic molecules. The specificity of the amplified DNA was confirmed by Southern blot analysis using the peroxidaselabelled probe and by the lack of amplified product with scallop DNA as the starting template.

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Section: Discussionsupporting

confidence: 67%

Genomic DNA cloning of rikettsia-like organisms (RLO) of Saint-Jacques scallop Pecten maximus: evaluation of pro-karyote diagnosis by hybridization with a non-isotopically labelled probe and by polymerase chain reaction

Kellner-Cousin¹,

Gall²,

Despres³

et al. 1993

Dis. Aquat. Org.

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“…1 8 Used in conjunction with DNA hybridisation, PCR can also detect single chlamydial organisms without the need for live organisms. 15,19…”

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confidence: 99%

Methods of detection of Chlamydia psittaci in domesticated and wild birds

McElnea¹,

Cross

1999

Aust Veterinary J

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“…The MOMP gene probe was generated by amplifying a fragment of approximately 1 kb from both C. psittaci and C. trachomatis omp1 genes (25). The groesl probe was generated by amplifying a fragment of approximately 730 bp from the C. psittaci and C. trachomatis groesl operons, using primers HS2 (5Ј-CCGCAT GCTCAGATCAAGCAACGACC-3Ј) and HS3 [5Ј-GGGAATTCATC(AG)A CAAC(AG)AC(CT)TT-3Ј].…”

Section: Methodsmentioning

confidence: 99%

Cytotoxic-T-lymphocyte-mediated cytolysis of L cells persistently infected with Chlamydia spp

1996

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Persistent chlamydial infections have been proposed as a means whereby chlamydiae evade immune resolution of infection. Such a mechanism would require evasion not only of the humoral immune responses but also of cell-mediated immune responses. We hypothesized that if such a mechanism is important, persistently infected cells should not be recognized by cytotoxic T cells. Persistent infections were simulated in vitro by treatment of Chlamydia trachomatis-or Chlamydia psittaci-infected cells with gamma interferon (IFN-␥), penicillin, or tryptophan depletion. Cultures were examined for induction of a chlamydial stress response (measured by transcription of groesl RNA) and for the effects on viability, infectivity, morphology, and immune recognition. Although both IFN-␥ and penicillin induced aberrant chlamydial morphology and growth, we did not find evidence that these treatments elicited a classical stress response. In addition, T-cell-mediated lysis of Chlamydia-infected target cells treated with IFN-␥ or penicillin or grown in tryptophan-deficient media was examined. The immune cell-mediated lysis of these treated infected cells demonstrated that despite the effects of these compounds on chlamydial growth and development, the infected cells continued to be efficiently recognized and killed by cytotoxic T cells. Thus, it seems unlikely that these in vitro models of persistence represent functional mechanisms to evade immune clearance.

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Detection of Chlamydia psittaci using DNA probes and the polymerase chain reaction

Cited by 6 publications

References 9 publications

Genomic DNA cloning of rikettsia-like organisms (RLO) of Saint-Jacques scallop Pecten maximus: evaluation of pro-karyote diagnosis by hybridization with a non-isotopically labelled probe and by polymerase chain reaction

Genomic DNA cloning of rikettsia-like organisms (RLO) of Saint-Jacques scallop Pecten maximus: evaluation of pro-karyote diagnosis by hybridization with a non-isotopically labelled probe and by polymerase chain reaction

Methods of detection of Chlamydia psittaci in domesticated and wild birds

Cytotoxic-T-lymphocyte-mediated cytolysis of L cells persistently infected with Chlamydia spp

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