Molecular Biology & Genomics Centre

Pathogen Identification


Molecular based methods like PCR, Real-Time PCR and more recently next generation sequencing (NGS) have revolutionized the field of veterinary diagnostics. At MBG, we offer detection of pathogenic viruses, bacteria and parasites using molecular methods, which are fast and highly sensitive to detect microbial pathogens in various specimens. MBG is an ISO 17025 accredited facility and benefits from an isolated containment level 3 facility for processing highly (level 3) contagious pathogens.


All Bacteria Virus Fungi Parasite


Accredited *
Assay Code See Below
Description African horse sickness (AHS) virus is a double stranded RNA virus of the family Reoviridae and causes African horse sickness in horses and related species such as mules, donkeys, and zebras. Horses are most severely affected by the disease. AHS is endemic in the central tropical regions of Africa and has occasionally extended to Egypt, the Middle East and the southern Arabian Peninsula.

The disease spreads mainly by the bite of insects such as midges (Culicoides) but direct horse to horse transmission does not occur. The virus can only survive through continuous cycles of transmission between its hosts-horses and insects. It does not survive in the environment outside of the host. Clinical signs are typically seen five to seven days after infection in the form of fever, redness of the inside surface of the eyelids. The disease progresses to one of the following forms: pulmonary form, cardiac form or mixed form.

There are nine distinct serotypes of AHSV (AHSV-1 to AHSV-9) . Any of the strains can cause disease with severity ranging from a mild fever to sudden death. The serotypes can be distinguished in serum neutralisation tests by the specificity of their reactions with neutralising antibodies. Molecular methods for determination of AHSV serotypes have been developed based on a set of nine individual ‘typing’ assays to detect and identify Seg-2 of each AHSV serotype.

Pathogens Tested
  • APA-001 : African horse sickness virus*
  • APA-182 : African horse sickness virus Serotyping

* Accredited Tests.

Method Real -Time RT-PCR
Sample Type
Accredited : EDTA Blood, Tissue (Spleen, Lung, Lymph nodes) , Culture.
Transport Condition Samples should be transported at 4°C and delivered within 24h of collection
Turn Around Time (TAT) Normal Turnaround time for pathogen Identification is within 3 working days.
Urgent Samples will be reported within half of the minimum test period & will be Charged Double.
Samples delivered after 11:00 AM will be processed next working day unless urgent.

Links

Validated
Assay Code APA-214
Description The genus Anaplasma belonging to the Anaplasmataceae family (order Rickettsiales) comprises obligate intracellular Gram-negative bacteria is one of the predominant tick borne pathogens of veterinary and public health importance. 
Anaplasma exhibit a biological cycle involving infection of both invertebrate and vertebrate host with different cell types targeted mainly infection of the red blood cells. Ticks transmit pathogens that cause disease thorough the process of feeding and the incubation period is 5-14 days. Affecting species are ruminants, dogs, horses, camels and also in human (Human granulocytic anaplasmosis). General systemic signs include anaemia, diarrhea, anorexia, and weight loss.
In the initial phase of infection, hosts are usually seronegative and serological cross-reactions with several Anaplasma species are observed. So, in terms of sensitivity and specificity PCR tests are the main approach for a case definition and epidemiological studies compared to other laboratory test methods.
In MBG, instead of detecting closely related species, we have been designed, a genus-specific primers to amplify a conserved region that target entire Anaplasma genus. One of the reasons is that some Anaplasma species are ecologically divergent and not found in the same hosts or vectors. Diagnosis for sensitive and specific identification of Anaplasma infections usually performed on total DNA from different kind of sample types including blood, culture but in persistently infected animals with intermittent or low-level bacteremia, other tissues might be useful.

Method PCR & Gel Electrophoresis
Sample Type
EDTA Blood
Transport Condition Sample should be transported at 4oC. (Refer to MBG-C0014)
Turn Around Time (TAT) Normal Turnaround time for pathogen Identification is within 3 working days.
Urgent Samples will be reported within half of the minimum test period & will be Charged Double.
Samples delivered after 11:00 AM will be processed next working day unless urgent.
Note Research use only.

Links

Validated
Assay Code See Below
Description Aspergillus spp. has been found worldwide in almost all domestic and wild animals, ranging from insects and corals to NHP. Aspergillosis is a common infection in birds, particularly pet parrots, penguins, captive raptors, mallards and other ducks, turkeys, and Japanese quail, in which it causes pulmonary and air sac infection. Aspergillosis has also been described in cats and dogs in which it affects only the nasal passages. In large animals, Aspergillus infection is assumed to be rare but has been reported with increasing frequency. It can lead to various diseases like mycotic abortion and gland infection in cows as well as guttural pouch involvement in horses. In marine mammals, airways are usually initially affected leading to pneumonia, but other organs including the brain may also be infected following fatal dissemination. The most frequently identified pathogens are Aspergillus fumigatus and Aspergillus flavus, ubiquitous organisms capable of living under extensive environmental stress.

Aspergillus fumigatus is an opportunistic pathogen and the primary cause of invasive aspergillosis. Triazole antifungals are used to treat aspergillosis. However, triazole-resistant A. fumigatus infections are increasingly reported worldwide and are associated with increased treatment failure and mortality. This fungus can acquire resistance to azole antifungals due to mutations in the azole target (Cyp51A). Cyp51A mutations typical for environmental azole resistance acquisition (for example, TR34/L98H) have been reported in many cases. These mutations can also be found in isolates recovered from patients.

Pathogens Tested
  • APA-098 : Aspergillus fumigatus/ terreus/ flavus
  • APA-189 : Azole-resistance profiling of Cyp51A gene

Method Aspergillus fumigatus / terreus / flavus by Multi Real-Time PCR (This assay includes the detection and differentiation of Aspergillus fumigatus / terreus / flavus)
Azole-resistance profiling of Cyp51A gene (This assay detects selected mutations in the Cyp51A gene related to azole-resistance, by PCR and sequencing).
Sample Type
Culture, EDTA Blood, Swab / Secretion (Respiratory), Tissue.
Transport Condition Sample should be transported at 4°C.
Turn Around Time (TAT) Normal Turnaround time for pathogen Identification is within 3 working days.
Urgent Samples will be reported within half of the minimum test period & will be Charged Double.
Samples delivered after 11:00 AM will be processed next working day unless urgent.

Links

Validated
Assay Code See Below
Description Aspergillus is a type of fungus that is found indoors and outdoors. Most people breathe in Aspergillus spores every day without getting sick. However, health problems may arise in people with weakened immune systems or having underlying diseases. Aspergillosis, most often caused by Aspergillus fumigatus, occurs in humans in chronic or acute forms which are clinically very distinct. Most cases of acute aspergillosis occur in people with severely compromised immune systems, e.g. those undergoing bone marrow transplantation. Chronic colonization or infection can cause complications in people with underlying respiratory illnesses, such as asthma, cystic fibrosis, sarcoidosis, tuberculosis, or chronic obstructive pulmonary disease. The most frequently identified pathogens are Aspergillus fumigatus and Aspergillus flavus, ubiquitous organisms capable of living under extensive environmental stress.

Aspergillus fumigatus is the primary cause of invasive aspergillosis. Triazole antifungals are used to treat patients with high-risk conditions such as stem cell and organ transplant recipients as mortality exceeds 50%. However, triazole-resistant A. fumigatus infections frequently occur and are associated with increased treatment failure and mortality. Of particular concern are resistant A.fumigatus isolates carrying either TR34/L98H or TR46/Y121F/T289A genetic resistance markers in the azole target (Cyp51A) gene and these are mostly associated with triazole fungicide use. Reports of these triazole-resistant A.fumigatus strains are more common in Europe than in the US.Understanding the prevalence of azole resistant patient isolates is important to guide clinical and public health decision-making.

Pathogens Tested
  • HPA-189 : Azole-resistance profiling of Cyp51A gene

Method Aspergillus fumigatus / terreus / flavus by Multi Real-Time PCR (This assay includes the detection and differentiation of Aspergillus fumigatus / terreus / flavus)
Azole-resistance profiling of Cyp51A gene (This assay detects selected mutations in the Cyp51A gene related to azole-resistance, by PCR and sequencing).
Sample Type
Accredited : 1
Alternatives : Culture, EDTA Blood, Swab / Secretion (Respiratory), Tissue.
Transport Condition Sample should be transported at 4°C.
Turn Around Time (TAT) TAT for routine samples is within 10 working days. Urgent Samples will be charged double and will be reported within 7 working days.
Samples delivered before 11:00 AM will begin processing immediately resulting in shorter TAT.
Note 555

Links

Validated
Assay Code APA-002
Description The adenoviruses are a group of non-enveloped, icosahedral DNA viruses. Adenovirus infections are ubiquitous in commercially farmed birds, and probably in all avian species.

The avian adenoviruses can be divided into three groups. Group I, or conventional adenoviruses, share a common group antigen. Group II adenoviruses include the viruses of turkey haemorrhagic enteritis (THE), marble spleen disease (MSD) and group II splenomegaly of chickens. These viruses also share a common antigen. Group III viruses, the egg drop syndrome (EDS) viruses, are widely distributed in waterfowl but can easily infect chickens, resulting in the production of abnormal eggshells. While many infections are subclinical and appear to be of little economic or welfare importance, significant outbreaks of disease associated with adenovirus do occur.

Method PCR and gel electrophoresis
Sample Type
Stool, Tissue (trachea, bursa, nasal mucosa, pharynx, lungs, kidney), Culture, Swab/Secretion (Rectal)
Transport Condition Samples should be transported at 4°C and delivered within 24h of collection.
Turn Around Time (TAT) Normal Turnaround time for pathogen Identification is within 3 working days.
Urgent Samples will be reported within half of the minimum test period & will be Charged Double.
Samples delivered after 11:00 AM will be processed next working day unless urgent.

Links




Molecular Biology & Genomics Centre

Pathogen Identification


Molecular based methods like PCR, Real-Time PCR and more recently next generation sequencing (NGS) have revolutionized the field of veterinary diagnostics. At MBG, we offer detection of pathogenic viruses, bacteria and parasites using molecular methods, which are fast and highly sensitive to detect microbial pathogens in various specimens. MBG is an ISO 17025 accredited facility and benefits from an isolated containment level 3 facility for processing highly (level 3) contagious pathogens.


All Bacteria Virus Fungi Parasite


Accredited *
Assay Code See Below
Description African horse sickness (AHS) virus is a double stranded RNA virus of the family Reoviridae and causes African horse sickness in horses and related species such as mules, donkeys, and zebras. Horses are most severely affected by the disease. AHS is endemic in the central tropical regions of Africa and has occasionally extended to Egypt, the Middle East and the southern Arabian Peninsula.

The disease spreads mainly by the bite of insects such as midges (Culicoides) but direct horse to horse transmission does not occur. The virus can only survive through continuous cycles of transmission between its hosts-horses and insects. It does not survive in the environment outside of the host. Clinical signs are typically seen five to seven days after infection in the form of fever, redness of the inside surface of the eyelids. The disease progresses to one of the following forms: pulmonary form, cardiac form or mixed form.

There are nine distinct serotypes of AHSV (AHSV-1 to AHSV-9) . Any of the strains can cause disease with severity ranging from a mild fever to sudden death. The serotypes can be distinguished in serum neutralisation tests by the specificity of their reactions with neutralising antibodies. Molecular methods for determination of AHSV serotypes have been developed based on a set of nine individual ‘typing’ assays to detect and identify Seg-2 of each AHSV serotype.

Pathogens Tested
  • APA-001 : African horse sickness virus*
  • APA-182 : African horse sickness virus Serotyping

* Accredited Tests.

Method Real -Time RT-PCR
Sample Type
Accredited : EDTA Blood, Tissue (Spleen, Lung, Lymph nodes) , Culture.
Transport Condition Samples should be transported at 4°C and delivered within 24h of collection
Turn Around Time (TAT) Normal Turnaround time for pathogen Identification is within 3 working days.
Urgent Samples will be reported within half of the minimum test period & will be Charged Double.
Samples delivered after 11:00 AM will be processed next working day unless urgent.

Links

Validated
Assay Code APA-214
Description The genus Anaplasma belonging to the Anaplasmataceae family (order Rickettsiales) comprises obligate intracellular Gram-negative bacteria is one of the predominant tick borne pathogens of veterinary and public health importance. 
Anaplasma exhibit a biological cycle involving infection of both invertebrate and vertebrate host with different cell types targeted mainly infection of the red blood cells. Ticks transmit pathogens that cause disease thorough the process of feeding and the incubation period is 5-14 days. Affecting species are ruminants, dogs, horses, camels and also in human (Human granulocytic anaplasmosis). General systemic signs include anaemia, diarrhea, anorexia, and weight loss.
In the initial phase of infection, hosts are usually seronegative and serological cross-reactions with several Anaplasma species are observed. So, in terms of sensitivity and specificity PCR tests are the main approach for a case definition and epidemiological studies compared to other laboratory test methods.
In MBG, instead of detecting closely related species, we have been designed, a genus-specific primers to amplify a conserved region that target entire Anaplasma genus. One of the reasons is that some Anaplasma species are ecologically divergent and not found in the same hosts or vectors. Diagnosis for sensitive and specific identification of Anaplasma infections usually performed on total DNA from different kind of sample types including blood, culture but in persistently infected animals with intermittent or low-level bacteremia, other tissues might be useful.

Method PCR & Gel Electrophoresis
Sample Type
EDTA Blood
Transport Condition Sample should be transported at 4oC. (Refer to MBG-C0014)
Turn Around Time (TAT) Normal Turnaround time for pathogen Identification is within 3 working days.
Urgent Samples will be reported within half of the minimum test period & will be Charged Double.
Samples delivered after 11:00 AM will be processed next working day unless urgent.
Note Research use only.

Links

Validated
Assay Code See Below
Description Aspergillus spp. has been found worldwide in almost all domestic and wild animals, ranging from insects and corals to NHP. Aspergillosis is a common infection in birds, particularly pet parrots, penguins, captive raptors, mallards and other ducks, turkeys, and Japanese quail, in which it causes pulmonary and air sac infection. Aspergillosis has also been described in cats and dogs in which it affects only the nasal passages. In large animals, Aspergillus infection is assumed to be rare but has been reported with increasing frequency. It can lead to various diseases like mycotic abortion and gland infection in cows as well as guttural pouch involvement in horses. In marine mammals, airways are usually initially affected leading to pneumonia, but other organs including the brain may also be infected following fatal dissemination. The most frequently identified pathogens are Aspergillus fumigatus and Aspergillus flavus, ubiquitous organisms capable of living under extensive environmental stress.

Aspergillus fumigatus is an opportunistic pathogen and the primary cause of invasive aspergillosis. Triazole antifungals are used to treat aspergillosis. However, triazole-resistant A. fumigatus infections are increasingly reported worldwide and are associated with increased treatment failure and mortality. This fungus can acquire resistance to azole antifungals due to mutations in the azole target (Cyp51A). Cyp51A mutations typical for environmental azole resistance acquisition (for example, TR34/L98H) have been reported in many cases. These mutations can also be found in isolates recovered from patients.

Pathogens Tested
  • APA-098 : Aspergillus fumigatus/ terreus/ flavus
  • APA-189 : Azole-resistance profiling of Cyp51A gene

Method Aspergillus fumigatus / terreus / flavus by Multi Real-Time PCR (This assay includes the detection and differentiation of Aspergillus fumigatus / terreus / flavus)
Azole-resistance profiling of Cyp51A gene (This assay detects selected mutations in the Cyp51A gene related to azole-resistance, by PCR and sequencing).
Sample Type
Culture, EDTA Blood, Swab / Secretion (Respiratory), Tissue.
Transport Condition Sample should be transported at 4°C.
Turn Around Time (TAT) Normal Turnaround time for pathogen Identification is within 3 working days.
Urgent Samples will be reported within half of the minimum test period & will be Charged Double.
Samples delivered after 11:00 AM will be processed next working day unless urgent.

Links

Validated
Assay Code See Below
Description Aspergillus is a type of fungus that is found indoors and outdoors. Most people breathe in Aspergillus spores every day without getting sick. However, health problems may arise in people with weakened immune systems or having underlying diseases. Aspergillosis, most often caused by Aspergillus fumigatus, occurs in humans in chronic or acute forms which are clinically very distinct. Most cases of acute aspergillosis occur in people with severely compromised immune systems, e.g. those undergoing bone marrow transplantation. Chronic colonization or infection can cause complications in people with underlying respiratory illnesses, such as asthma, cystic fibrosis, sarcoidosis, tuberculosis, or chronic obstructive pulmonary disease. The most frequently identified pathogens are Aspergillus fumigatus and Aspergillus flavus, ubiquitous organisms capable of living under extensive environmental stress.

Aspergillus fumigatus is the primary cause of invasive aspergillosis. Triazole antifungals are used to treat patients with high-risk conditions such as stem cell and organ transplant recipients as mortality exceeds 50%. However, triazole-resistant A. fumigatus infections frequently occur and are associated with increased treatment failure and mortality. Of particular concern are resistant A.fumigatus isolates carrying either TR34/L98H or TR46/Y121F/T289A genetic resistance markers in the azole target (Cyp51A) gene and these are mostly associated with triazole fungicide use. Reports of these triazole-resistant A.fumigatus strains are more common in Europe than in the US.Understanding the prevalence of azole resistant patient isolates is important to guide clinical and public health decision-making.

Pathogens Tested
  • HPA-189 : Azole-resistance profiling of Cyp51A gene

Method Aspergillus fumigatus / terreus / flavus by Multi Real-Time PCR (This assay includes the detection and differentiation of Aspergillus fumigatus / terreus / flavus)
Azole-resistance profiling of Cyp51A gene (This assay detects selected mutations in the Cyp51A gene related to azole-resistance, by PCR and sequencing).
Sample Type
Accredited : 1
Alternatives : Culture, EDTA Blood, Swab / Secretion (Respiratory), Tissue.
Transport Condition Sample should be transported at 4°C.
Turn Around Time (TAT) TAT for routine samples is within 10 working days. Urgent Samples will be charged double and will be reported within 7 working days.
Samples delivered before 11:00 AM will begin processing immediately resulting in shorter TAT.
Note 555

Links

Validated
Assay Code APA-002
Description The adenoviruses are a group of non-enveloped, icosahedral DNA viruses. Adenovirus infections are ubiquitous in commercially farmed birds, and probably in all avian species.

The avian adenoviruses can be divided into three groups. Group I, or conventional adenoviruses, share a common group antigen. Group II adenoviruses include the viruses of turkey haemorrhagic enteritis (THE), marble spleen disease (MSD) and group II splenomegaly of chickens. These viruses also share a common antigen. Group III viruses, the egg drop syndrome (EDS) viruses, are widely distributed in waterfowl but can easily infect chickens, resulting in the production of abnormal eggshells. While many infections are subclinical and appear to be of little economic or welfare importance, significant outbreaks of disease associated with adenovirus do occur.

Method PCR and gel electrophoresis
Sample Type
Stool, Tissue (trachea, bursa, nasal mucosa, pharynx, lungs, kidney), Culture, Swab/Secretion (Rectal)
Transport Condition Samples should be transported at 4°C and delivered within 24h of collection.
Turn Around Time (TAT) Normal Turnaround time for pathogen Identification is within 3 working days.
Urgent Samples will be reported within half of the minimum test period & will be Charged Double.
Samples delivered after 11:00 AM will be processed next working day unless urgent.

Links