Commercial Diagnostic Laboratory Research Articles

Clinical, histological and prognostic features of a novel nail-bed lesion of cats: 41 cases.

Objectives There is a distinct subset of lesions arising on the digits of cats, located at or close to the nail-bed epithelium, which are typically composed of proliferative fibroblast-like cells, multinucleate giant cells and areas of osseous metaplasia, but currently there is no published literature detailing the clinical or histological features of these lesions. Methods This study identified 41 such cases from two large commercial diagnostic laboratories and assessed various histological and clinical features; 22 cases had additional follow-up data available. Results All masses in this study were exophytic, variably inflamed, contained large numbers of spindle cells and had areas of capillary formation. The majority also had areas of ulceration, multinucleate giant cells and osseous metaplasia. The mitotic count was variable, but mitoses were confined to the fibroblast-like cells. Male cats appeared predisposed and the second digit was the most commonly affected. Conclusions and relevance These distinctive lesions arising on the digits of cats had potential for local recurrence but metastasis was not reported. Based on these clinical and histological features, the masses in this study appear most similar to giant cell reparative granulomas, and trauma, injury to the nail or nail-bed and nail-bed infections may potentially contribute to their development.

Plasma concentrations of coffee polyphenols and plasma biomarkers of diabetes risk in healthy Japanese women.

Coffee consumption has been reported to reduce the risk of type 2 diabetes in experimental and epidemiological studies. This anti-diabetic effect of coffee may be attributed to its high content in polyphenols especially caffeic acid and chlorogenic acid. However, the association between plasma coffee polyphenols and diabetic risks has never been investigated in the literature. In this study, fasting plasma samples were collected from 57 generally healthy females aged 38–73 (mean 52, s.d. 8) years recruited in Himeji, Japan. The concentrations of plasma coffee polyphenols were determined by liquid chromatography coupled with mass tandem spectrometer. Diabetes biomarkers in the plasma/serum samples were analysed by a commercial diagnostic laboratory. Statistical associations were assessed using Spearman's correlation coefficients. The results showed that plasma chlorogenic acid exhibited negative associations with fasting blood glucose, glycated hemoglobin and C-reactive protein, whereas plasma total coffee polyphenol and plasma caffeic acid were weakly associated with these biomarkers. Our preliminary data support previous findings that coffee polyphenols have anti-diabetic effects but further replications with large samples of both genders are recommended.

Abstract PD7-02: Multiplex Identification of genetic etiologies among women with bilateral breast cancer using a 25-gene hereditary cancer panel

Abstract Background: Multiple primary cancers within an individual is one of the hallmarks of hereditary cancer predisposition. Technical advances in sequencing and identification of additional cancer susceptibility genes have led to multi-gene panel approaches to determine if patient cancers have a heritable cause. Simultaneous testing of multiple breast cancer associated genes to determine the prevalence, spectrum and combinations of mutations has not yet been evaluated in a large set of patients with two breast cancers. Methods: Patients with two breast cancer diagnoses were identified from 80,748 consecutive cases that underwent a 25-gene hereditary cancer panel test at a commercial diagnostic laboratory. Patient clinical data were obtained by healthcare provider report on test requisition forms. Results: Of 3,182 patients with two breast cancers, 13.3 % (n=424) had a pathogenic variant (PV) in at least one of 19 genes on the panel. PVs were most prevalent within BRCA1, representing 26.3% of the PVs, followed by BRCA2 (21.9%), CHEK2 (13.5%), PALB2 (11%) and ATM (11%). Fourteen patients had PVs in two different genes, including five with CHEK2/PALB2 in combination, which was the only combination observed more than once. PVs were more common among the 33.8% of patients with synchronous breast cancers compared to those with metachronous diagnoses (10.8% vs. 15%) and in the 58.4% who were first diagnosed younger than 50 years old (15.7% vs. 10%). Conclusion: Multiplex testing in women with two primary breast cancers identifies a relatively high percentage with a PV, including those whose first diagnosis was older than 50. While still the most prevalent and clear cause, fewer than half of the PVs identified were in BRCA1 or BRCA2. The observation of 5 cases with double CHEK2/PALB2 PVs is statistically unexpected given their individual prevalence in other studies of high risk women with breast cancer, suggesting a possible synergistic or additive effect. This study adds to our understanding of susceptibility to multiple primary breast cancers, and reaffirms the importance of this circumstance as a prompt for genetic testing. Nonetheless, as 66% of cases had metachronous tumors, it would be preferable to identify those at risk with heritable cancer syndromes at their first diagnosis and enable subsequent prevention or early detection. Citation Format: Weitzel JN, Blazer KR, Nehoray B, Kidd J, Slavin Jr TP, Solomon I, Niell-Swiller M, Rybak C, Saam J, Lancaster J. Multiplex Identification of genetic etiologies among women with bilateral breast cancer using a 25-gene hereditary cancer panel. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr PD7-02.

Abstract PD7-03: Characterization of Li-Fraumeni syndrome diagnosed using a 25-gene hereditary cancer panel

Abstract Background: Clinical diagnostic criteria for Li-Fraumeni syndrome (LFS) have evolved with increased utilization of TP53 germline testing and subsequent improved understanding of the diversity of the associated cancer phenotypes. However, data on LFS still suffer from ascertainment bias as patients are typically selected to undergo TP53 testing based on the presence of hallmark features of LFS. Analyzing TP53 mutation carriers identified from multi-gene panel testing, for which the diagnosis of LFS may not have been suspected or was included in a longer differential diagnosis, affords an opportunity to characterize additional TP53 carriers who might not otherwise have been ascertained. Methods: Patients with a deleterious or suspected deleterious germline TP53 mutation were identified from 80,748 consecutive cases that underwent a 25-gene hereditary cancer panel test between September 2013 and March 2015 at a commercial diagnostic laboratory. Patient clinical data were obtained by healthcare provider report on test requisition forms. Each TP53 mutation carrier was evaluated to determine whether the National Comprehensive Cancer Network's (NCCN) guidelines were met for TP53 testing. Results: Eighty-one TP53 mutation carriers were identified and had a total of 115 cancers (0.1% overall prevalence). Among the 76 carriers with at least one cancer, the average age at first diagnosis was 42 years (range 11-76 years) and 24% were first diagnosed older than age 50 years. The most common first cancers were of the breast (n=45), ovary (n=9), and gastrointestinal tract (n=8). Fifty-two of the 75 (69%) women had breast cancer, 44% of which were first diagnosed at 35 years or younger, and 21% were first diagnosed at 50 years or older. Only 27 TP53 carriers met NCCN criteria for TP53 testing, 14 of whom only met based on having early-onset breast cancer. An additional 8 did not meet criteria themselves but had a first- or second-degree relative who did. Among the 28 individuals with more than one primary cancer, 21 (75%) developed their second primary at a site for which increased surveillance is recommended in LFS, but only 4 would have met NCCN criteria for TP53 testing at their first cancer diagnosis. The most common second cancers were of the breast (n=16), gastrointestinal tract (n=4), or kidney (n=2) and occurred an average of 11 years after the first cancer (range 0-36 years). Conclusion: In this analysis, a large proportion of carriers would not have been identified as TP53 testing candidates based on NCCN guidelines. Our data are consistent with other studies demonstrating high second primary cancer risks in LFS, and highlight the value of multigene panel testing in identifying individuals who may be candidates for increased surveillance and/or cancer risk-reducing management options. Citation Format: Rich T, Lotito M, Kidd J, Saam J, Lancaster J. Characterization of Li-Fraumeni syndrome diagnosed using a 25-gene hereditary cancer panel. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr PD7-03.

Elevated plasma and urinary concentrations of green tea catechins associated with improved plasma lipid profile in healthy Japanese women

This study investigated green tea catechins in plasma and urine and chronic disease biomarkers. We hypothesized that plasma and urinary concentration of green tea catechins are associated with cardiovascular disease and diabetes biomarkers. First void urine and fasting plasma samples were collected from 57 generally healthy females aged 38 to 73 years (mean, 52 ± 8 years) recruited in Himeji, Japan. The concentrations of plasma and urinary green tea catechins were determined by liquid chromatography coupled with mass tandem spectrometer. Low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, triglyceride, glucose, insulin, glycated hemoglobin, and C-reactive protein in plasma/serum samples were analyzed by a commercial diagnostic laboratory. Statistical associations were assessed using Spearman correlation coefficients. The results showed weak associations between plasma total catechin and triglyceride (r = −0.30) and LDL cholesterol (r = −0.28), whereas plasma (−)-epigallocatechin-3-gallate, (−)-epigallocatechin, (−)-epicatechin-3-gallate, and (−)-epicatechin exhibited weak to moderate associations with triglyceride or LDL cholesterol, but little associations with HDL cholesterol, body fat, and body mass index were evident. Urinary total catechin was weakly associated with triglyceride (r = −0.19) and LDL cholesterol (r = −0.15), whereas urinary (−)-epigallocatechin-3-gallate (r = −0.33), (−)-epigallocatechin (r = −0.23), and (−)-epicatechin-3-gallate (r = −0.33) had weak to moderate correlations with triglyceride and similarly with body fat and body mass index. Both plasma (r = −0.24) and urinary (r = −0.24) total catechin, as well as individual catechins, were weakly associated with glycated hemoglobin. Plasma total and individual catechins were weakly to moderately associated with C-reactive protein, but not the case for urinary catechins. In conclusion, we found weak to moderate associations between plasma and urinary green tea catechin concentrations and plasma biomarkers of cardiovascular disease and diabetes.

Abstract P4-12-02: Spectrum of mutations identified in a 25-gene hereditary cancer panel for patients with breast cancer

Abstract Introduction: With the advancements of next generation sequencing, patients with a personal and/or family history of cancer that may not be suggestive of one cancer syndrome may be offered testing for mutations in multiple cancer-predisposing genes simultaneously. The focus of this analysis was to determine the spectrum of gene mutations observed in patients with a personal history of breast cancer. Methods: A commercial diagnostic laboratory database was queried for patients with a personal diagnosis of breast cancer who underwent a 25-gene hereditary cancer panel from September 4, 2013 through April 17, 2014. The panel includes highly penetrant cancer predisposing genes BRCA1, BRCA2, TP53, PTEN, MLH1, MSH2, MSH6, PMS2, EPCAM, APC, BMPR1A, CDH1, CDKN2A, MUTYH, SMAD4, STK11 and moderately penetrant genes CHEK2, PALB2, ATM, NBN, BARD1, BRIP1, CDK4, RAD51C and RAD51D. Sequencing and large rearrangement was performed for all the genes in the panel. All patient data regarding clinical history was obtained by health care provider report on the test requisition forms. Results: A total of 3584 patients with a personal history of breast cancer were identified. Of these, 13.8% met the NCCN guidelines for genetic testing for both Hereditary Breast and Ovarian Cancer syndrome (HBOC) and Lynch syndrome (LS), while 80.3% met criteria for only HBOC and 1.2% met only for LS. 10.4% of females and 18.9% of males with breast cancer were positive for at least 1 deleterious or suspected deleterious mutation, of which mutations in BRCA1 and BRCA2 comprised 40.2%. In this cohort, 59.8% of the mutations were detected in other genes (see table below). Of the patients who did not meet either testing criteria, mutations were found in BRCA2 (2), APC (2), BARD1 (1), CHEK2 (1), MSH2 (1), NBN (1), and TP53 (1). Interestingly, we also identified 15 patients with two mutations in our cohort. BRCA1 or BRCA2 accounted for one of the mutations in 9 out of 15 patients and 6 patients had two mutations in other genes. Gene% of Total MutationsBRCA121.2%BRCA219.0%CHEK211.3%ATM9.7%PALB27.2%NBN6.6%APC5.6%BARD13.3%PMS22.6%BRIP12.3%MSH62.0%TP531.5%MSH21.3%RAD51C1.3%CDH11.0%RAD51D1.0%CDKN2A1.0%PTEN1.0%MLH10.5%Biallelic MUTYH0.3%SMAD40.3% Conclusions: Testing patients using a 25-gene panel identified 234 mutations outside of BRCA 1 and BRCA 2 (157). That is a 149% increase in mutations identified over BRCA 1 and BRCA 2 testing alone. Mutations in moderately penetrant breast cancer genes (including CHEK2, ATM, PALB2 and NBN) comprised 34.8% of total mutations and 6.4% of mutations were in LS genes. Additionally, panel testing identified mutations in more than one gene in 4.0% of patients, which would not have been identified by single-syndrome testing. Panel testing provides a broader understanding of hereditary cancer in breast cancer patients both by identifying mutations in more genes and identifying patients with mutation in more than one gene. This approach can provide more guidance both for management of the patient and the patient's family members. Citation Format: Lavania Sharma, Kelsey Moyes, John Abernethy, Heidi McCoy, Jennifer Saam, Michelle Landon, Richard Wenstrup. Spectrum of mutations identified in a 25-gene hereditary cancer panel for patients with breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-12-02.

Discovery of Heterodera filipjevi in Washington and Comparative Virulence with H. avenae on Wheat

The cereal cyst nematode Heterodera avenae suppresses wheat production in the western United States. A second species of cereal cyst nematode, H. filipjevi, was identified in eastern Oregon during 2008. This paper reports the discovery of H. filipjevi-infested fields in eastern Washington, thereby extending the known distribution of H. filipjevi in the United States. The identity of H. filipjevi was determined and confirmed by species-specific polymerase chain reaction (PCR), PCR-restriction fragment length polymorphism (RFLP), sequencing, and cyst morphology. Soils that were collected from naturally infested fields in Washington were used to compare the virulence of H. avenae and H. filipjevi on six spring wheat cultivars under controlled-environment conditions. Noninfested soils from nearby fields were used as controls. Cultivars Ouyen and WB Rockland were resistant to H. avenae and susceptible to H. filipjevi. Cultivars Sönmez and SY Steelhead were resistant to H. filipjevi and susceptible to H. avenae. Cultivars Louise and WB 936 were susceptible to both species. The resistance of SY Steelhead to 'H. avenae', reported in a previous paper, is corrected as resistance to H. filipjevi due to an earlier misidentification of H. filipjevi. Management guidelines that include crop rotations and resistant cultivars are presented. Discovery of additional infestations of H. filipjevi are anticipated when DNA-based tests become used routinely in commercial diagnostic laboratories.

Interlaboratory and between-specimen comparisons of diagnostic tests for leptospirosis in sheep and cattle

A study was performed to investigate interlaboratory test agreement between a research and a commercial veterinary diagnostic laboratory on blood and urine samples, and to investigate test agreement between blood, urine, and kidney samples (research laboratory) for leptospirosis diagnosis. Samples were sourced from 399 sheep and 146 beef cattle from a local abattoir. Interlaboratory agreement for real-time quantitative polymerase chain reaction (qPCR) results on urine samples was almost perfect (kappa = 0.90), despite the use of different amplification targets (DNA gyrase subunit B gene vs. 16s ribosomal RNA gene), chemistries (SYTO9 vs. TaqMan probe), and pre-PCR processing. Interlaboratory agreement for microscopic agglutination test (MAT) positivity was almost perfect (kappa = 0.93) for Leptospira borgpetersenii serovar Hardjo subtype Hardjobovis (Hardjobovis) but moderate (kappa = 0.53) for Leptospira interrogans serovar Pomona (Pomona). Among animals that had different titers recorded, higher Hardjobovis and lower Pomona titers were reported by the commercial laboratory than by the research laboratory (P < 0.005). These interlaboratory comparisons can assist researchers and diagnosticians in interpreting the sometimes discrepant test results. Within the research laboratory, the comparison of qPCR results on urine and kidney showed almost perfect agreement (kappa = 0.84), suggesting that the qPCR on these 2 specimens can be used interchangeably. The agreement between MAT positivity and urine and kidney qPCR results was fair (kappa = 0.32 and kappa = 0.33, respectively). However, the prevalence ratio of urine and kidney qPCR positivity in Hardjobovis-seropositive versus Hardjobovis-seronegative sheep indicated that Hardjobovis seropositivity found in sheep may be able to predict shedding or renal carriage.

Writing diagnostic laboratory requisition form histories

JAVMA, Vol 244, No. 4, February 15, 2014 V submit samples to diagnostic laboratories for a wide variety of reasons, including routine wellness testing, preanesthetic testing, and, most notably, diagnostic testing for disease. A necessary component of sample submission is inclusion of a laboratory requisition form, which serves to communicate important clinical information about the patient so that the laboratory diagnostician has a reasonable context for interpreting results during sample analysis. However, a common observation among laboratory diagnosticians at university-based and commercial diagnostic laboratories is that many requisition forms either provide no clinical information or provide information that is so incomplete or disorganized as to be of little use. There are no published statistics on the prevalence of this problem, but anecdotally, this observation is commonly advanced in the circles of board-certified pathologists and clinical pathologists, in both human and veterinary medicine. History and clinical context are fundamental considerations in interpreting diagnostic test results, with many pathologists and clinical pathologists likely to agree that sample interpretation is perhaps 50% to 80% context. This may be even higher for small samples, such as cytologic, endoscopic biopsy, and needle biopsy samples, for which sample quality may be limited. History is also important when concerns arise that samples may have been mixed either at the practice prior to submission or at the diagnostic laboratory itself. A good clinical history aids the laboratory diagnostician in confirming the integrity of the sample being examined. Given the importance of providing a complete history when submitting samples for diagnostic testing, veterinarians should consider using a template to ensure that all critical information is included on the requisition form. By following a few practical suggestions and answering 6 questions regarding the animal and sample, veterinarians can ensure that they are providing the context required for an informed interpretation of any type of sample by any qualified individual.

Rubella immunity in Japanese women of childbearing age, 2008-2013: commercial diagnostic laboratory data analysis

For effective non-routine vaccination in women of childbearing age as a countermeasure against congenital rubella syndrome (CRS), it is important to know the specific age group in which fewer members have rubella immunity. We analyzed rubella hemagglutination-inhibition (HI) antibody data (about 890,000) accumulated from 2008 through 2013 at a commercial diagnostic laboratory, which originated from the serum specimens sent mainly from obstetrics & gynecology clinics and hospital departments in Japan. Changes during the above period in the pattern of the annual curve for the prevalence of rubella antibodies (HI antibody titer > or = 8) by age, were observed. The antibody prevalence among women in their twenties decreased gradually from 2008 to 2013. However, the prevalence at < or = 22 years of age in the 2013 specimens was found to have risen, which we believe is the effect of the five-year interim vaccination program for high-school students that began in 2008. The lowest antibody prevalence at 23 years of age in 2013 corresponded to the highest numbers of female patients at the same age in the 2013 rubella epidemic among the adult population. The analysis of extensive data accumulated for rubella antibodies assayed at the commercial diagnostic laboratory was useful for monitoring the susceptibility of the women to rubella infection in Japan, and that the Oct. 1987 to Mar. 1990 birth cohort (age 23 - 25 in 2013) is a high-risk group for CRS.

Comparative Cost-Effectiveness of Two-Tiered Testing Strategies for Serodiagnosis of Lyme Disease with Noncutaneous Manifestations

The mainstay of laboratory diagnosis for Lyme disease is two-tiered serological testing, in which a reactive first-tier enzyme-linked immunosorbent assay (ELISA) or an immunofluorescence assay is supplemented by separate IgM and IgG immunoblots. Recent data suggest that the C6 ELISA can be substituted for immunoblots without a reduction in either sensitivity or specificity. In this study, the costs of 4 different two-tiered testing strategies for Lyme disease were compared using the median charges for these tests at 6 commercial diagnostic laboratories in 2012. The study found that a whole-cell sonicate ELISA followed by the C6 ELISA was the most cost-effective two-tiered testing strategy for Lyme disease with acute-phase serum samples. We conclude that the C6 ELISA can substitute for immunoblots in the two-tiered testing protocol for Lyme disease without a loss of sensitivity or specificity and is less expensive.

Sensitivity of a real-time polymerase chain reaction for Tritrichomonas fetus in direct individual and pooled preputial samples

The objective of this study was to evaluate the sensitivity of a commercially available real-time polymerase chain reaction (PCR) test for the detection of Tritrichomonas fetus in individual and pooled direct preputial samples. Two samples were collected and processed once a week from nine T. fetus–infected bulls (n = 121) and placed into either an InPouch TF or 2 mL of PBS. Preputial samples were also collected into both media and PBS from 1016 other bulls. All pouches were cultured and evaluated as per manufacturer's instructions. The prepuce samples collected directly into PBS were individually evaluated using real-time PCR by a commercial diagnostic laboratory. Direct preputial samples were then randomly divided for pooling into groups of 5 and 10 samples, ensuring that every pool had one sample from a known infected bull before testing using real-time PCR. Sensitivity was estimated for culture and real-time PCR of the 121 direct and culture-enriched individual samples from nine infected bulls. There were no differences (P = 0.12) among the sensitivity estimates for culture, 95.0% (95% confidence interval [CI]: 89.6%–97.7%); real-time PCR of culture-enriched samples, 95.9% (95% CI: 90.7–98.2); and direct preputial samples, 90.1% (95% CI: 83.5–94.2). There was also no significant difference (P = 0.08) between the sensitivity of real-time PCR for direct preputial samples in 110 pools of 5 (83.6%, 95% CI: 75.6–89.4) or 10 samples (77.3%, 95% CI: 68.6–84.1). The use of three sequential direct samples, collected in PBS at weekly intervals and tested by real-time PCR, increased the sensitivity to 100% for pools of 5 and 96% for pools of 10. In conclusion, direct preputial samples collected in PBS and tested by real-time PCR individually have comparable sensitivity to culture and real-time PCR in enriched samples. The use of pooled direct preputial samples appears to be relatively sensitive. However, this strategy requires repeated sampling to optimize sensitivity. Real-time PCR testing of preputial samples collected directly into PBS with the option of pooling would decrease the cost associated with screening bulls, and increase the feasibility of large epidemiological studies and active surveillance.

Development of a broad-range quantitative polymerase chain reaction assay to detect and identify fungal DNA in equine endometrial samples

To develop a broad-range 28S ribosomal DNA quantitative PCR (qPCR) assay for detection of fungal DNA in equine endometrial samples. 12 fungal samples from a clinical diagnostic laboratory and 29 samples obtained from 17 mares. The qPCR assay was optimized with commercially acquired fungal organisms and validated with samples obtained from the clinical diagnostic laboratory. Subsequently, 29 samples from 17 mares suspected of having fungal endometritis were evaluated via the qPCR assay and via traditional fungal culture and endometrial cytology. Amplicons from the qPCR assay were subjected to genetic sequencing to identify the organisms. The qPCR assay theoretically had a detection threshold of 2 organisms of Candida albicans. Fungal DNA was amplified from all 12 fungal samples from the commercial diagnostic laboratory. Fungal identification by use of genetic sequencing was successful for 34 of 36 amplicons from the 12 samples assayed. A fungal agent was identified via qPCR assay and genetic sequencing in all 12 samples; in contrast, a fungal agent was identified in only 8 of 12 samples via standard fungal culture and biochemical analysis. The qPCR assay detected fungal DNA in samples from 12 of 17 mares suspected of having fungal endometritis. A rapid, sensitive, and repeatable qPCR assay was developed for detection of fungal DNA from equine endometrial samples. The qPCR may prove to be clinically useful as an adjunct to microbial culture and cytologic examination to provide identification of fungal organisms in a timely manner.

Antimicrobial Resistance and Pharmacodynamics of Canine and Feline Pathogenic E. coli in the United States

Percent resistance and minimum inhibitory concentrations (MIC) were described for canine (n = 301) and feline (n = 75) pathogenic Escherichia coli (E. coli) isolates solicited during May 2005 to Sep 2005 from the Clinical Pharmacology Laboratory at Auburn University (n = 165) or commercial diagnostic laboratories ([CDL]; n = 211) from four regions in the USA. Drugs tested were amoxicillin (AMX), amoxicillin trihydrate/clavulanate potassium (AMXC), cefpodoxime (CFP), doxycycline (DXY), enrofloxacin (ENR), gentamicin (GM) and trimethoprim-sulfamethoxazole (TMS). Urinary isolates were most common (n = 174). Percent resistance was greatest for isolates from the respiratory tract, urine, and skin compared with the ear. Resistance was also greatest for samples sent from the south and central states compared with the western states (P ≤ 0.001). Percent resistance by drug was AMX (46 ± 2.6%) > AMXC (37 ± 2.5%) > CFP (21.8 ± 2%) = DXY (22 ± 2.1%) = ENR (20 ± 2.1%) = TMS (19 ± 2%) > GM (12 ± 1.7%). There was a significant difference in resistance between the different antibiotic drugs (P ≤ 0.001). Population MIC distributions were bimodal, and MICs were highest in samples from the southern states (P ≤ 0.001). E. coli resistance may limit its empirical treatment. For susceptible isolates, AMX and AMXC may be least effective and TMS most effective.

On a Chip

The future of clinical and POC BioMEMS is very bright. With an increasing emphasis on the personalization of medicine and the rising costs of health care, early detection and diagnostics at the POC will be even more important. Early detection implies early intervention, resulting in the saving of lives and reducing overall spending. The potential impact of these technologies on the early diagnosis and management of both communicable and noncommunicable diseases is very high. Many grand challenges applications are possible, e.g., routine tests such as complete blood cell count on a chip that an individual can perform at home; detection of cardiac markers from blood after a perceived heart attack; detection of cancer markers such as exosomes, CTCs from blood, or protein biomarkers in serum; and detection of infectious agents such as virus and bacteria for public health. These applications are expected to result in new diagnostic assays for home, doctor's office, clinical laboratories, and various POC settings.

A descriptive analysis of the antimicrobial susceptibility of mastitis-causing bacteria isolated from samples submitted to commercial diagnostic laboratories in New Zealand (2003–2006)

AIM: To describe the antimicrobial susceptibility of mastitis-causing bacteria isolated from milk samples submitted to commercial laboratories over a period of 40 months. METHODS: The records of reported results of milk samples submitted by veterinary practitioners to five commercial veterinary laboratories in the North and South Islands of New Zealand, between August 2003 and December 2006 were reviewed. Logistic regression was used to analyse the effect of year, island, and the interaction of year and antimicrobial on the probability of antimicrobial susceptibility for each pathogen and antimicrobial combination, where the causative bacteria had >1,000 susceptibility tests in total and the antimicrobials was tested on >500 isolates. A total of 9,262 isolates were included in this study, with an average of nearly seven susceptibility tests per isolate, totalling 62,918 tests. RESULTS: Streptococcus uberis isolates demonstrated high overall susceptibility (>90.0%) to the majority of antimicrobial agents except ampicillin (81.7%), lincomycin (85.3%), trimethoprim/sulphonamide combination (88.6%), and, as expected, aminoglycosides (<4%). The susceptibility of Strep. dysgalactiae was similar to that of Strep. uberis, except for greater susceptibility to oxacillin (98.1%) and much lower susceptibility to tetracyclines (11.2%). The susceptibility of Staphylococcus aureus isolates was markedly different from that of Strep. uberis for the majority of antimicrobials tested. Susceptibility of Staph. aureus was lower than 90% to ampicillin (73.4%), erythromycin (74.7%), lincomycin (66.1%), penicillin (73.1%), and streptomycin (71.7%). No antimicrobial was effective against all Staph. aureus isolates. Minor changes were found in the overall susceptibility of the main mastitis-causing bacteria between 2003 and 2006. CONCLUSIONS: The antimicrobial agents intended for treatment of bovine mastitis currently available in New Zealand generally demonstrated good in-vitro efficacy against streptococci and staphylococci, with the exception of aminoglycosides. CLINICAL RELEVANCE: Analysis of the results of antimicrobial susceptibility of bacterial isolates from milk samples from dairy cows in New Zealand provides useful data for surveillance purposes, and a baseline for identifying changes in antimicrobial sensitivity in this population. However, the variation in antimicrobial susceptibility between individual isolates means that these data are of limited value when determining treatment of mastitis at the farm level.

Canine paediatric oncology: retrospective assessment of 9522 tumours in dogs up to 12 months (1993-2008)

Little information is available on the occurrence of neoplasms in dogs up to the age of 12 months. This is a retrospective review of histopathological diagnoses of neoplasia in dogs up to the age of 12 months based on biopsy specimens submitted to a commercial veterinary diagnostic laboratory in the United Kingdom between 1993 and 2008. In 20 280 histological submissions, 9522 neoplasms were identified. Canine cutaneous histiocytoma (n = 8465; 89%) was the most common histological type. Neoplasms other than histiocytoma (n = 1057; 11%) were grouped as benign epithelial (n = 375; 4%), haematopoietic (n = 229; 2%), benign mesenchymal (n = 145; 2%), miscellaneous (n = 118; 1%), non-hematopoietic malignant mesenchymal (n = 118; 1%) or malignant epithelial tumours (n = 72; <1%). Excluding canine cutaneous histiocytoma, 52% of tumours (n = 547) were benign, and 66% were from the skin or soft tissues. These data provide valuable epidemiological information on neoplasms occurring in juvenile dogs in the United Kingdom.

Laboratory Testing Practices for West Nile Virus in the United States

We surveyed state public health and commercial diagnostic reference laboratories regarding current testing practices for West Nile virus (WNV). The majority of WNV testing is now performed in commercial diagnostic reference laboratories using commercially available Food and Drug Administration-cleared kits labeled for the presumptive diagnosis of WNV. However, only 25% of surveyed state public health or commercial diagnostic reference laboratories currently have the capacity to perform the recommended confirmatory testing. These findings indicate the need for both manufacturers and laboratories to monitor the performance of these WNV test kits. Further, clinicians should be aware of the limitations of these kits and the need for additional testing to confirm a diagnosis of WNV disease.

Detection of antibodies against Leptospira serovars via microscopic agglutination tests in dogs in the United States, 2000–2007

To use results of microscopic agglutination tests (MATs) conducted at a commercial veterinary diagnostic laboratory to determine temporal and demographic distributions of positive serologic test results for leptospirosis in dogs and identify correlations among results for various Leptospira serovars. Serosurvey. MAT results for 33,119 canine serum samples submitted to a commercial veterinary diagnostic laboratory from 2000 through 2007. Electronic records of MAT results for dogs were obtained from a veterinary diagnostic laboratory. Seropositivity for antibodies against Leptospira serovars was determined by use of a cutoff titer of >or=1:1,600 to reduce the possible impact of postvaccinal antibodies on results. Correlations between results for all possible pairs of serovars were calculated by ordinal ranking of positive (>or=1:100) antibody titer results. 2,680 samples (8.1%; 95% confidence interval [CI], 7.8% to 8.4%) were seropositive for antibodies against Leptospira serovars. The highest percentage of positive MAT results was for the year 2007 (10.2%; 95% CI, 9.5% to 10.9%) and for the months of November and December during the study period. Antibodies were most common against serovars Autumnalis, Grippotyphosa, Pomona, and Bratislava. Seroprevalence of leptospirosis was lowest for dogs>10 years of age but was similar across other age strata. Leptospirosis can affect dogs of small and large breeds and various ages. Although an increase in proportions of positive MAT results was evident in the fall, monthly and annual variations suggested potential exposure in all months. Because of the limitations of MAT results and the limited number of serovars used in the test, bacterial culture should be used to identify infective Leptospira serovars.

Contemporary prevalence of infectious agents in laboratory mice and rats

Periodic health screening of rodents used in research is necessary due to the consequences of unwanted infections. One determinant of the risk of infection for any given agent is its prevalence; other factors being equal, a prevalent agent is more likely than a rare one to be introduced to a research facility and result in infection. As an indicator of contemporary prevalence in laboratory populations of rats and mice, the rate of positive results in the samples received at a major commercial rodent diagnostic laboratory was compiled for this paper. Although samples from laboratory rodent vendors have been excluded, results are tabulated from samples from more than 500,000 mice and 80,000 rats submitted over several years from pharmaceutical, biotechnology, academic, and governmental institutions in North America and Europe, allowing meaningful determination of which agents are common in the research environment versus which agents are rare. In mice, commonly detected infectious agents include mouse norovirus, the parvoviruses, mouse hepatitis virus, rotavirus, Theiler's murine encephalomyelitis virus, Helicobacter spp., Pasteurella pneumotropica, and pinworms. In rats, commonly detected infectious agents include 'rat respiratory virus', the parvoviruses, rat theilovirus, Helicobacter spp., P. pneumotropica, and pinworms. A risk-based allocation of health-monitoring resources should concentrate frequency and/or sample size on these high-risk agents, and monitor less frequently for the remaining, lower-risk, infectious agents.