PCRmax Chlamydophila felis Instruction Manual

Major outer membrane protein
(ompA gene)

Chlamydophila felis

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

1

Chlamydophila felis is a rod shaped Gram negative bacterium that is part of the
Chlamydiaceae family and responsible for infections of the eye in cats. C. felis is an
obligately intracellular, parasitic pathogen with a rigid, lipid containing cell wall. It is
predominantly isolated from cats but also seen in humans. The genome is of circular
confirmation and around 1.16 mbp long, containing 1005 protein-coding genes. It also
contains a 7.5 kbp circular plasmid that is required for virulence. Other virulence factors
include the major outer membrane proteins and polymorphic outer membrane proteins that
are required for host cell entry.

The bacterium cannot survive outside of the host and is transmitted through contact with
ocular secretions from infected individuals. C. felis attacks the conjunctival epithelium of
the eye, entering cells after attachment to sialic acid receptors. The organism cannot
replicate autonomously and has an unusual replication cycle that occurs in the cytoplasm
of the epithelial cells, culminating in formation of the infectious form. Bacterial induced lysis
of the cells leads to release of the infectious bodies and spread of infection to other
epithelia throughout the body.

Clinical signs include watery discharge from the eye, chemosis, red and swollen
conjunctivae, mild nasal discharge, sneezing, lethargy, pneumonitis and may cause
infertility in cats. In young kittens chlamydiosis may cause pneumonia. The incubation
period is 2-5 days and the infection is common in cats aged less than a year old. Clinical
signs usually appear in one eye and progress to become bilateral. Conjunctival shedding
usually ceases at around 60 days after infection but persistent infections may develop.

Introduction to Chlamydophila felis

2

MIN

MAX

The PCR Max qPCR Kit for Chlamydophila felis (C.felis) genomes is designed for the in
vitro quantification of C.felis genomes. The kit is designed to have the broadest detection
profile possible whilst remaining specific to the C.felis genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
C.felis sequences based on a comprehensive bioinformatics analysis.

If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to [e-mail] and our bioinformatics team will
answer your question.

Specificity

3

Kit Contents

• C.felis specific primer/probe mix (150 reactions BROWN)

FAM labelled

• C.felis positive control template (for Standard curve RED)

• Internal extraction control primer/probe mix (150 reactions BROWN)

VIC labelled as standard

• Internal extraction control DNA (150 reactions BLUE)

• Endogenous control primer/probe mix (150 reactions BROWN)

FAM labelled

• RNAse/DNAse free water (WHITE)

for resuspension of primer/probe mixes and internal extraction control DNA

• Template preparation buffer (YELLOW)

for resuspension of positive control template and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.

Lyophilised 2x qPCR Mastermix

This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.

Pipettors and Tips

Vortex and centrifuge

Thin walled 1.5 ml PCR reaction tubes

4