PCRmax Enterobacter cloacae complex Instruction Manual

Transcriptional activator (ramA)
gene

Enterobacter cloacae
complex

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Enterobacter cloacae is a Gram-negative, facultatively-anaerobic, rod-shaped bacterium
that is part of the normal gut flora. However, they can be responsible for various infections,
including bacteremia, lower respiratory tract infections, skin and soft-tissue infections,
urinary tract infections (UTIs), endocarditis, intra-abdominal infections, septic arthritis,
osteomyelitis, CNS, and ophthalmic infections. The DNA genome is typically about 5.5Mb
and consists of a single circular chromosome with plasmids present depending on the
strain. Genes encoding adhesion and invasion proteins are typically found on the
chromosome along with iron chelating and hemolysin-like proteins. The size of this
bacteria ranges from 0.3-0.6 x 0.8-2.0 μm. Enterobacter cloacae lives in a mesophilic
environment with its optimal temperature at 37 °C and uses its peritrichous flagella for
movement. This organism is oxidase negative but catalase positive and can make ATP by
aerobic respiration when oxygen is present but can switch to fermentation in the absence
of oxygen.

Enterobacter cloacae are nosocomial pathogens that can cause a range of infections as
previously mentioned. Due to their prevalence in the body, this organism affects mostly
the vulnerable age groups such as the elderly and the young and can cause prolonged
hospitalization. During infection, microbes displaying NulO sugar mimicry may
downregulate host complement-mediated killing and be advantageous in a wide range of
animal body habitats. The organism carries genes for 37 multidrug efflux proteins, 7
antimicrobial peptide resistance proteins, and 11 β-lactamases thereby making treatment
extremely difficult.

Enterobacter cloacae infections that target the bladder and urinary tract may cause pain or
an uncomfortable pressure. Many people suffering from a urinary infection with
Enterobacter cloacae complain of a frequent urge to urinate, pain or burning while
urinating, and reduced urine flow. In the respiratory system, the bacterium causes
breathing problems and pneumonia. Patients with respiratory Enterobacter cloacae suffer
from shortness of breath, yellow sputum (phlegm), fevers and heavy coughing.
Interestingly, pneumonia caused by this bacterium often makes patients feel less ill than
pneumonia caused by other bacteria, but has a surprisingly high mortality rate.
Enterobacter cloacae may also infect the inner surface of the heart valves, or endocardia,
causing new heart murmurs, weight loss, coughing, and fevers. People with this type of
infection may feel extremely tired, especially when they exercise. They may also have
bloody urine, and problems associated with insufficient blood circulation. When the
bacterium enters a joint, it can produce a form of arthritis. This is most likely to happen if
the bacteria is already in the blood stream and is carried to the joint, or if it enters the body
from an injury near the joint. Symptoms are sudden, and include severe pain as the
bacteria attack the joint, swelling, fever and chills. A subtype of E.cloacae, Enterobacter
cloacae subsp. Dissolvens has been investigated as a potential producer of high yield
biofuels.

Introduction to Enterobacter cloacae complex

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MAX

MIN

The Primerdesign™ genesig® Kit for the Enterobacter cloacae complex (E.
cloacae_complex) is designed for the in vitro quantification of members of the E.cloacae
complex. The kit is designed to have the broadest detection profile possible whilst
remaining specific to the E.cloacae complex.

This kit will detect:

Enterobacter cloacae, Enterobacter asburiae, Enterobacter hormaechei, Enterobacter
kobei and Enterobacter ludwigii.

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.

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

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Kit Contents

• E.cloacae_complex specific primer/probe mix (150 reactions BROWN)

FAM labelled

• E.cloacae_complex 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

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