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Newsletter 10/2017
We are pleased to welcome you to the monthly BattLab newsletter. This newsletter will bring you the latest news and information about our laboratory and all tests that we can offer to all our clients.
From our whole staff, a great big thank you to all the veterinarians that last month attended our fourth (and last for 2017) seminar on diagnostic approach to anaemia in cats. We hope you found the talk inspirational and we look forward to seeing you at the next seminars we are in the process of planning for 2018. Keep an eye on our Website and Facebook page for more information.
FAQ about PCR-based diagnostic for infectious diseases
What is PCR?
PCR is a diagnostic molecular technique that has recently become very popular in veterinary medicine. PCR amplifies a single copy (or a few copies) of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. This technique is applicable to several areas of veterinary medicine, from the identification of infectious diseases to genetic based diseases.
Why choose PCR for the identification of infectious diseases?
PCR, similarly to culture but different from serology, is able to identify the agent itself rather than the immune response to it, which can take up to several weeks before becoming apparent. It is also useful for the detection of agents for which culture is difficult or it requires specialist sampling (eg. Mycobacterium, Leptospira) and it does not require the presence of viable organisms.
Which are the recommended samples for PCR testing?
The type of sample required for PCR testing varies from test to test and the pathogenesis of the condition under investigation. PCR is a very sensitive technique, but correct sample selection is still essential, since a negative result indicates the absence of the pathogen in the collected sample, but the infection may still be present elsewhere in the body. Samples that can be used for PCR testing include:
  • EDTA blood samples: this is the preferred sample for organisms that have tropism for cells in the blood stream (e.g. Mycoplasma). EDTA is preferred over lithium heparin, since the latter can inhibit the PCR reaction. 
  • Swabs: only dry swabs without transport medium.
  • Fresh faeces: for selected organisms (e.g. Tritrichomonas foetus) that are intermittently shed, a pooled faecal is preferred 
  • Urine and fluids
  • Cytology samples: PCR can also performed on prestained slides and does not require additional sampling.
  • Tissue samples: fresh tissue samples in saline are preferred over formalin fixed ones since the formalin fixation may crosslink and fragment DNA reducing the sensitivity of the test.
Samples can be sent through regular post and do not require special treatment.
Which pathogens can be tested with PCR?
A large variety of infectious diseases affecting small animals can be tested with PCR, from haemotropic Mycoplasma, to Feline Calicivirus, Feline Coronavirus, FeLV, FIV, Toxoplasma and many more. We recommend checking our catalogue which includes an extensive list of the PCR tests offered, including interesting combined PCR profiles at a special price.
Which are the most common causes of false negative results in PCR testing?
Negative results may be observed when the target nucleic acid does not exceed the detectable limit in that sample or may not be present at all in the sample, despite its presence elsewhere in the body. This is why a correct selection of the sample to submit is so crucial.
Urolithiasis is a general term referring to the causes and effects of calculi in the urinary tract. Under less than optimal conditions, some wastes, especially minerals, precipitate out of solution to form crystals. If these crystallized minerals are retained in the urinary system, they may grow and aggregate to form calculi. 
Risk factors that are known to affect canine uroliths include:
  • Breed: Oxalate uroliths are most common in Miniature Schnauzer and urate uroliths in Dalmatians.
  • Gender: Females dogs are predisposed for infection-induced struvite uroliths.
  • Age: urolithiasis is more common in middle-aged/adult dogs.
  • Anatomical and functional abnormalities of the urinary tract.
  • Abnormalities of metabolism: ammonium urate uroliths are often associated with portosystemic shunt (PSS).
  • Urinary tract infections: increased risk for infection-induced struvite uroliths is noted in case of  UTI caused by E. coli and S. aureus.
  • Diet.
  • Urine pH: an alkaline urine pH is a risk factor for struvite and calcium phosphate uroliths, while calcium oxalate, urate, cystine and silica uroliths are more commonly observed in acidic urine.
Clinical signs associated with urolithiasis depend on the location of the stone and its size.  Formation of macroscopic uroliths in the lower urinary tract that interfere with the flow of urine and/or irritate the mucosal surface often results in dysuria, haematuria, and stranguria. When present in the upper parts of the urinary tract, clinical signs may vary from absent to more severe.
Diagnosis. Multiple uroliths may be present throughout the urinary tract, therefore a complete radiographic examination of the tract is indicated; radiodense calculi >3 mm in diameter are usually visible on radiographs. Urate, and occasionally cystine, uroliths may be radiolucent, requiring contrast radiography or ultrasonography to confirm their presence. 
Urinalysis, including identification of crystals on microscopic examination is a critical part of the evaluation and may help determine the type of urolith present. However, the absence of crystals on urinalysis, does not exclude the possibility of uroliths in the urinary tract.
Knowing the urolith composition is important, because this has a major impact on the treatment and prevention, therefore calculi analysis after removal is always strongly recommended.
Calculi analysis At Battlab, the analysis of dog uroliths is performed by Fourier Transform Infrared Spectroscopy (FTIR). FTIR is a quantitative technique that shows a high sensitivity and allows an accurate identification of stone composition; it is now becoming the gold standard for stone analysis. The infrared spectrum originates from the vibrational motion of the molecules, which are a kind of fingerprint of the compounds. This property is used for characterization of organic and inorganic compounds present in the calculi. The band intensities are proportional to the compound concentration and hence qualitative and quantitative estimations are also obtained. One mineral usually predominates, but the composition of many uroliths can be also mixed. Our internal data of the samples analysed in the last two years showed that most of the submitted stones were struvite uroliths (56%), followed by calcium oxalate (22%), cystine (7.6%) and ammonium urate (7.6%). 
Sample requirements for uroliths are as following; they must be submitted dry, in their entirety, not placed in formalin, in non-crushable mailing containers. All the retrieved stones must be sent, as they may have different mineral layers. 
Turnaround time is 2-3 working days only. 
For more information and prices please contact our laboratory. 
Veterinary Surgeon and Company Director at Broad Lane Vets in Coventry
There has been a Veterinary Practice on Broad Lane since 1969; it was first established by Roy Hands, and his son Roger later followed in his footsteps. I graduated from Liverpool Vet School and then worked at the PDSA in Aston, Birmingham, before joining Broad Lane Vets as an assistant Veterinary Surgeon in 2001. I like a challenge, so when Roger stepped down, I took on the running of the practice, and I'm still enjoying mixing vetting with management nearly 10 years later.
Growing up, I always took the class hamster or gerbil home for the holidays, and even the stick insects. As a family, we always had goldfish and cats, and even a chinchilla, and eventually I persuaded my parents to get a dog, to keep me company on my newspaper round. I volunteered on Saturdays at Birmingham Nature Centre as soon as I was old enough, and then at a local vet practice. In common with many of my fellow vets, I suspect, I always preferred animals to people - of course you later realise that every pet comes with a person attached! Over the years I have learned to embrace the interaction with clients, and I do get tremendous satisfaction out of working in partnership with a pet owner, for the common aim of improving their pet's health and wellbeing. However, you'd never call me a natural people-person!
We've been using Battlab for laboratory services since Roger Batt started the business, many years ago now. We have built up a relationship with their team and have picked the Battlab vets' brains many times! The improved accuracy of their results, compared to those of a small in-house machine, is unquestionable. However, we particularly like the personal friendly touch that we get from Battlab, and the fact that we can speak to a person there easily, without sitting on hold for hours. On occasion, Battlab have even phoned and personally alerted us to a result that really concerned them, which has meant we have been able to get that pet in to re-check it or start it on potentially life-saving treatment at the earliest possible opportunity. That impressed us. With the lab being so geographically close, we're also able to drop-off additional samples before or after the daily Battlab collection. We look forward to a continued excellent working relationship with Battlab.
Yours sincerely,
The BattLab team
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