rodent and rabbit assay data sheet
- Ultrasensitive qualitative detection of
by real time polymerase chain reaction
- Ultrasensitive qualitative detection of
by real time polymerase chain reaction
- Ultrasensitive qualitative detection
of Helicobacter bilis
and Helicobacter hepaticus
by real time PCR.
is included on
P0032 Mouse Minipanel)
Helicobacter species screen by real time polymerase
chain reaction. This screen detects but
does not differentiate
H. pylori, H. heilmannii, H.
bilis, H. hepaticus,
H. rappini, H. felis, H. salomonis and other
Helicobacter species identification by real time
polymerase chain reaction and PCR product sequence analysis. This 2-stage assay detects
and differentiates H.
pylori, H. heilmannii, H. bilis, H. hepaticus,
H. rappini, H. felis, H. salomonis and
other Helicobacter species.
Many species of
the genus Helicobacter
have been identified in mammals and their pathogenicity varies.
Some species can induce significant disease while others appear
to merely colonize the gastrointestinal tract.
species that have been identified in mice and rats include
H. muridarum, H. rappini, H.
H. bilis, all of which colonize the gastrointestinal
tract. Among them, H.
muridarum and H.
rappini are generally not considered to be
pathogenic. However, in susceptible strains of mice
H. hepaticus causes
acute focal, nonsuppurative, necrotizing hepatitis which
progresses to chronic, active hepatitis characterized by minimal
necrosis, hepatocytomegaly, oval cell hyperplasia and
cholangitis. Typically, relatively few infectious organisms are
apparent on routine histologic examination. Mouse strains A/JCr,
B6C3F1, BALB/cAnCr, C3H/HeNCr, SJL/NCr, and SCID/NCr have been
shown to develop hepatitis due to
infection, and infected A/JCr and male B6C3F1 mice show
increased incidence of hepatocellular neoplasms.
can be found in the bile, liver, and intestines of mice.
Infection of both rats and mice with this bacterium is
associated with chronic hepatitis. It is not known whether
H. bilis causes
liver tumors in mice. However, infection of rat or mouse liver
by H. hepaticus
can significantly affect the interpretation of carcinogenicity
bioassays and other research using rodents.
In the past,
definitive diagnosis of H.
hepaticus- or H.
bilis-associated liver disease in mice was based on
the presence of histologic alterations as well as culture of the
infectious agent. However,
Helicobacter identification by culture from tissue
or feces is very difficult due to the organism’s fastidious
growth requirements. Sensitivity of the culture method is low.
organisms have been detected in lesioned liver by methods such
as Steiner and Warthin-Starry histochemical stains,
immunoflourescence, in situ hybridization, immunohistochemistry
and ultrastructural analysis. A serum ELISA test for circulating
IgG antibodies against H.
hepaticus has also been used to provide information
for determining in vivo infection status in pathogenesis
studies. However, none of these methods are very useful for
large-scale screening. PCR-based diagnostic assays for
H. hepaticus and
H. bilis offer
greater specificity, higher sensitivity and lower cost compared
to culture, histochemical stains, or ultrastructural analysis.
Recently, the growing use of real time PCR technology has
further improved sensitivity and specificity of molecular
detection of Helicobacter
species, making this method the most important approach for
routine colony screening. Real time PCR detection also allows
retrospective studies of paraffin-embedded tissue and other
archived tissue biopsies.
is a gram-negative spiral bacterium found in gastric mucosa and
associated with active and chronic gastritis.
H. pylori can
establish a chronic, persistent infection, which may lead to
gastric or duodenual ulcers, gastric cancer and gastric
lymphomas. Studies have revealed that approximately 50% of the
world’s human population is infected with
Biochemically, the bacterium produces catalase, oxidase and
urease enzymes. The urease enzyme permits the bacterium to
metabolize urea present in the gastric mucosa and establish a
microenvironment favorable to the organism.
H. pylori is a
highly motile organism with multiple unipolar flagella. Both the
urease enzyme and the flagella are considered to be important
infection in humans relies on upper endoscopy or the 13C-urea
breath test (see review by Nakamura, 2001). Although the
endoscopy procedure permits culture of the bacterium from biopsy
specimens (the gold standard for diagnosis), demonstration of
urease activity and histological detection of the germ, the
procedure is expensive and invasive. The 13C-urea breath test is
a well-established, relatively sensitive, specific and
noninvasive method. Molecular tests, such as PCR, can also offer
precise diagnosis of H.
pylori infections. In fact, molecular testing by PCR
can complement other diagnostic tests because it can be applied
to archival fixed tissue, environmental samples, gastric fluid,
oral secretions, and stool samples, in which traditional
diagnostic tests do not have sensitivity and perform poorly.
Studies have shown than PCR detection of
H. pylori in gastric
fluid specimens can reach a sensitivity of 96% and a specificity
of 100% (Westblom et al., 1993; Yoshida et al., 1999). This
capability is especially useful in monitoring active
H. pylori infection
in rodents and other animals, as the breath test is difficult to
conduct for these animals.
(previously known as
Gastrospirillum hominis) is a 4-10 µm long,
spiral-shaped, motile bacterium with three to eight coils, a
wavelength of about 1 µm, up to 14 uni- or bipolar flagella, and
no periplasmic filaments. In humans, gastric infection with
H. heilmannii is
associated with the development of chronic gastritis (found in
the stomachs of 0.2 to 4% of patients with gastritis) and
low-grade mucosa-associated lymphoid tissue lymphoma in humans.
Eradication of H. heilmannii
by antibiotic treatment of patients can result in
complete remission of MALT lymphoma, indicating a causal
relationship between H.
heilmannii infection and MALT lymphoma. Unlike
infections, gastric infections with
H. heilmannii or
Gastrospirillum-like organisms are not restricted to humans. A
broad range of animals including dogs, cats, pigs, and cattle
are naturally infected, with frequencies ranging from 80% to
100%. It has been suggested that
infection in humans may be a zoonosis and that animals may serve
as a reservoir for transmission to humans.
culture of H. heilmannii
has not been achieved to date (Anderson et al.,
1996) and diagnosis of H. heilmannii infection is usually made on the basis of its
distinct spiral morphology, compared with
H. pylori, on
silver- stained tissue sections. However, there are a number of
large, gastric, spiral organisms such as
H. felis, H. salomonis, and H. bizzozeronii which are
indistinguishable from H.
heilmannii on routine light microscopy, and
H. pylori grown in a
broth culture can also adopt a morphology identical to that of
(Fawcett et al., 1999). Molecular detection methods are required for more definitive identification (Trebesius et al.,
2001). PCR is the most sensitive and specific tool for detecting
infection (Whary and Fox, 2006).
Help confirm the disease causing agent
Shorten the time required to confirm a clinical
Help ensure that rodent colonies are free of
Early prevention of spread of these bacteria among a
Minimize personnel exposure to these bacteria
Safety monitoring of biological products that derive
1 fecal pellet,
or 0.2 ml fresh, frozen or fixed tissue. For
H. pylori and H. heilmannii
assays only, 0.2 ml gastric wash is the preferred
Contact Zoologix if advice is needed to determine an appropriate specimen type for a specific diagnostic application. For specimen types not listed here, please contact Zoologix to confirm specimen acceptability and shipping instructions.
specimen types, if there will be a delay in shipping, or during
very warm weather, refrigerate specimens until shipped and ship
with a cold pack unless more stringent shipping requirements are
specified. Frozen specimens should be shipped so as to remain
frozen in transit. See shipping
instructions for more information.
Whary, M.T. and Fox, J.G. (2006). Detection, eradication, and
research implications of Helicobacter infections in laboratory
rodents. LabAnimal, Vol. 35 No. 7:25-36.
2 business days (1 week for P0011)
B0021, B0023, B0039 and
- Qualitative real time PCR
Qualitative real time PCR + PCR product sequence analysis