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Evaluation and use of three cowside tests for detection of subclinical ketosis in early postpartum cows.

The objective was to evaluate the performance of 3 cowside diagnostic tests for detection of subclinical ketosis, defined as a serum beta-hydroxybutyrate (BHBA) concentration >or=1400 micromol/L. On 16 d over a 5-mo period, samples of serum, milk, and urine were collected on a large dairy facility from cows of all parities between 2 and 15 DIM. The sample proportion of subclinical ketosis was 7.6% (n = 859 samples from 545 cows). The KetoCheck powder (Great States Animal Health, St. Joseph, MO) detecting acetoacetate in milk samples was very specific (99%) but poorly sensitive (41%). Respective sensitivities and specificities of the Ketostix strip detecting acetoacetate in urine samples (Bayer Corporation, Elkhart, IN) were 78 and 96% with a cut-off point of "small", or 49 and 99% with a cut-off of "moderate." The KetoTest strip (Sanwa Kagaku Kenkyusho Co. Ltd., Nagoya, Japan) using milk samples had a sensitivity and specificity of 73 and 96% with a cut-off of 100 micromol of BHBA/L or 27 and 99% with a cut-off of 200 micromol of BHBA/L. On average, use of the Ketostix at the "small" cut-off point or the KetoTest at 100 micromol/L would result in no more than 3 or 4 false positives per 100 cows screened, with prevalence levels ranging from 5 to 30%, whereas the number of false negatives would range from one false negative at 5% prevalence to 7 or 8 false negatives at 30% prevalence. Either the Ketostix or KetoTest strips would provide acceptable results for screening individual cows on commercial dairies to detect subclinical ketosis. Over this prevalence range, the KetoCheck powder test would have limited application as a screening test. Despite only one false positive per 100 animals screened, false negatives resulting from screening with the KetoCheck test would be too frequent, ranging from 3 false negatives at 5% prevalence to 18 at 30% prevalence in a population of 100 tested cows. Finally, given their relative imprecision, use of any of these individual cowside tests to estimate herd prevalence must be done cautiously, especially when only a small number of animals are sampled.

3-Hydroxybutyric Acid↗

Visual tests for urinary amylase investigated in the routine laboratory.

Ninety-seven fresh urine specimens were tested in the routine laboratory with the Rapignost Amylase test strip, and the results were compared to those obtained using Phadebas Amylase test tablets to investigate the transferability of the results obtained by the Rapignost method to those of the Phadebas method under routine conditions. The fraction of conflicting negative results of Phadebas-positive specimens was 9% and the corresponding fraction of conflicting positive results was 13%. An attempt to improve the transferability by changing the comparison scale slightly did not succeed. However, a visual binary test based on 180 s incubation at 37 degrees C of 400 microliters urine in a suspension of a Phadebas Amylase test tablet seemed more suitable in selection of specimens with Phadebas amylase activity less than 2000 U/l (upper limit of the reference interval). These specimens amounted to approximately 75% of all amylase specimens in our laboratory.

Amylases↗

Reliability of crevicular fluid measurements taken in the presence of supragingival plaque.

The measurement of gingival crevicular fluid (GCF) is commonly used in clinical studies to quantitate the severity of gingival inflammation. This study examined the influence of the presence of supragingival plaque on GCF readings. Bilateral pairs of maxillary premolars received replicate GCF measurements. One of the teeth was randomly selected to be the test tooth. Supragingival plaque was removed from the test tooth between the first and second GCF measurements. Plaque was not removed from the control tooth. The crevicular fluid was collected on filter paper strips which were placed just subgingivally. The quantity of crevicular fluid was determined with the aid of the Periotron 6000. The mean difference between the first and second GCF measurement for the control teeth was -1.5 +/- 5.7 (+/- SD). This change was not significant (P = 0.21). The mean difference between the first and second GCF measurements for the experimental teeth (-21.3 +/- 16.5) was significant (P less than or equal to .001). The mean difference between the control and the experimental GCF differences were found to be highly significant (P less than or equal to .001). The data indicate that the presence of supragingival plaque can significantly elevate the GCF measurements when determined with the Periotron 6000.

Adult↗

Anaerobic bag culture method.

In a new method of anaerobic culture, a transparent, gas-impermeable bag is used and the anaerobic environment is established with copper sulfate-saturated steel wool. An Alka-Seltzer tablet generates carbon dioxide. The agar plate surface can be inspected through the bag at any time without interrupting the anaerobic atmosphere or disturbing other specimens. Methylene blue indicator strips are completely reduced by 4 h after the bag is set up and have remained reduced for as long as 3 weeks. Growth of 16 different stock culture anaerobes was generally equivalent by the bag and GasPak jar methods. Yield and growth of anaerobic isolates also were equivalent with 7 of 10 clinical specimens; from the other 3 specimens, 13 isolates were recovered, 5 by both the bag and jar methods and the rest by one method or the other. No consistent differences were found between the anaerobic bag and GasPak jar methods in the yield of anaerobes from clinical specimens. Early growth (24 h of incubation) of anaerobes from one specimen was detected with the bag method.

Anaerobiosis↗

Identification of Staphylococcus species with the API STAPH-IDENT system.

The API STAPH-IDENT system was compared with conventional methods for the identification of 14 Staphylococcus species. Conventional methods included the Kloos and Schleifer simplified scheme and DNA-DNA hybridization. The API STAPH-IDENT strip utilizes a battery of 10 miniaturized biochemical tests, including alkaline phosphatase, urease, beta-glucosidase, beta-glucuronidase, and beta-galactosidase activity, aerobic acid formation from D-(+)-mannose, D-mannitol, D-(+)-trehalose, and salicin, and utilization of arginine. Reactions of cultures were determined after 5 h of incubation at 35 degrees C. Results indicated a high degree of congruence (greater than 90%) between the expedient API system and conventional methods for most species. The addition of a test for novobiocin susceptibility to the API system increased the accuracy of identification of S. saprophyticus, S. cohnii, and S. hominis, significantly. Several strains of S. hominis, S. haemolyticus, and S. warneri which were difficult to separate with the Kloos and Schleifer simplified scheme were accurately resolved by the API system.

Bacteriological Techniques↗

Determining prevalence of bluetongue and epizootic hemorrhagic disease viruses in mule deer in Arizona (USA) using whole blood dried on paper strips compared to serum analyses.

We investigated the feasibility of using whole blood dried on paper strips as a means to collect antibody prevalence data for the epizootic hemorrhagic disease viruses (EHDV) and bluetongue viruses (BTV) from hunter-harvested male mule deer (Odocoileus hemionus) in October 2002 from Arizona, USA. We compared antibody prevalence estimates in mule deer from paired paper strip and serum samples. Prevalence data obtained from elution of dried blood on paper strips proved to be consistent with results from serum in 94% of the samples tested. The paper strip method allows easy collection of blood from dead animals, with a smaller amount of blood being needed for analyses. Also, samples do not need to be refrigerated before analyses. We also used serum samples to determine hemorrhagic disease (HD) serotype exposure status of mule deer harvested from 4 distinct areas in Arizona. Antibodies to BTV and EHDV were identified in 3 of the 4 areas, with positive results to EHDV-1, EHDV-2, BTV-10, and BTV-11 being most common. Many animals did not have antibodies against the BTV serotypes. Exposure varied geographically and potentially with elevation. Hemorrhagic disease viruses commonly infect Arizona mule deer, except on the Kaibab Plateau in northern Arizona.

Animals↗

Detection of immunoglobulin G to a Sindbis-related virus by a membrane antigen enzyme immunoassay.

We determined the seroprevalence of a Sindbis-related virus isolated for the first time in 1975 from ticks in south-east Sicily and typed by Gresikova et al. in 1978. An indirect enzyme immunoassay based on viral membrane antigen for coating microtiter strips was used for the detection of immunoglobulin G to the Sindbis-related virus. The method appeared more sensitive than a similar enzyme immunoassay based on crude lysate antigen. Comparison of the results obtained from sera tested both by membrane antigen enzyme immunoassay and microneutralization test showed 92% agreement, while the agreement between microneutralization test and crude antigen enzyme immunoassay was 76%. An overall elevated seroprevalence (63.66%) was found in a population group living in and around the area of first isolation and seroprevalence in different age groups was also studied.

Adolescent↗

Caries experience and streptococci and lactobacilli salivary levels in 6-8-year-old Sardinians.

OBJECTIVES: The aim of this investigation was to compare clinical data regarding oral status (DMFT/DMFS indices) and streptococci or lactobacilli counts in stimulated whole saliva in two groups of children, aged 6 and 8 years. METHODS: Seventy-nine boys and 93 girls aged 6 (84 subjects) and 8 (88 subjects) years old were selected. DMFT/dmft, DMFS/dmfs and OHI-S indices were recorded. Mutans streptococci (Sm) and lactobacilli (Lb) salivary levels were assessed using commercially available strip tests and were rated from 0 to 3 and from 0 to 4 in the group as a whole. RESULTS: Mean dmfs was 9.06 +/- 10.80, made up of ds = 7.28 +/- 9.54, fs = 1.19 +/- 3.64 and ms = 0.59 +/- 3.47. Mean DMFS was 0.73 +/- 1.91, made of DS = 0.54 +/- 1.29, FS = 0.08 +/- 0.56 and MS = 0.11 +/- 0.56. Statistically significant differences were found for both Sm and Lb scores between those with and without caries in the primary teeth (P < 0.05). The different Lb scores were also significant in relation to caries in permanent teeth in older children. CONCLUSION: A high proportion of children had some caries experience, and were in need of treatment.

Analysis of Variance↗

Evaluation of dipstrips, direct gram stain and pyuria as screening tests for the detection of bacteriuria.

Two hundred and fifty cases of clinically suspected urinary tract infection were analysed for the detection of bacteriuria. Parameters studied included direct Gram staining, pyuria on microscopic examination of uncentrifuged urine and dip strip method for the detection of blood, protein, nitrite and leucocyte esterase. Significant bacteriuria (colony count 10(5) per ml) was found in 112 cases with a positivity ranging from 65 to 83% for the presence of blood, protein, nitrite and leucocyte esterase. Highest positive predictive values were obtained with the presence of nitrite and leucocyte esterase (98%), blood, protein and nitrite (94%) as well as with blood, protein, nitrite and leucocyte esterase (98%). Both pyuria and direct Gram staining were positive in 85% cases. The combined presence of both these parameters gave 100% positive predictive value. Gram staining combined with pyuria was more effective and economical as compared to the dipstrips for the detection of bacteriuria.

Bacteriological Techniques↗

Mutans streptococci and dental caries prevalence in a group of Latvian preschool children.

Paraffin-stimulated saliva samples were collected from 140 children 3- and 4-yr old attending nine nursery schools in Latvia. The salivary levels of mutans streptococci were rated from zero to 3 after being cultured on a commercially available strip selective for these microorganisms. Of the children, 29.3% were rated at zero (approximately < 10(4) cfu per ml saliva). This group of children demonstrated the lowest mean caries prevalence dmfstot = 1.5 (SD 1.9). The highest dmfstot was found among children in class 2 (38.6%; approximately > 10(5)-10(6) cfu/ml) and class 3 (12.1%; approximately > 10(6) cfu/ml) with a mean caries prevalence of 6.5 (SD 5.8) and 6.4 (SD 6.0), respectively. The study demonstrates the association between high caries prevalence and high salivary levels of mutans streptococci in the young child. It is suggested that early identi-fication of mutans streptococci-colonized children might be of value in selecting at caries risk children for preventive measures.

Child, Preschool↗

Simplified screening for microalbuminuria.

BACKGROUND: Screening for microalbuminuria is increasingly advocated as a way to diagnose early renal involvement in diabetes and other diseases. It usually entails the use of a radioimmunoassay that is expensive and not always readily available. OBJECTIVE: To assess the efficacy of three simple and inexpensive tests for ruling out microalbuminuria. DESIGN: Cross-sectional study. SETTING: Outpatient clinics. PATIENTS: 221 patients from primary care clinics and a diabetes clinic. MEASUREMENTS: Random urine specimens were tested for albumin by using Micral-Test immunoassay strips (Boehringer Mannheim, Mannheim, Germany) and for protein by using sulfosalicylic acid testing and impregnated dipsticks (Chemstrips, Boehringer Mannheim). Radioimmunoassay for albumin was used for all specimens as standard for comparison. RESULTS: When less than 20 mg/L was considered the upper limit of normal for albumin concentration, Micral-Test, sulfosalicylic acid testing, and Chemstrips had negative predictive values of 99%, 95%, and 96%, respectively. Seventy-four specimens tested negative on both sulfosalicylic acid and Chemstrips; the negative predictive value of these two tests combined was 99%. CONCLUSIONS: The combination of sulfosalicylic acid testing and Chemistrips was as good as and less expensive than Micral-Test in ruling out microalbuminuria.

Albuminuria↗

Rapid urinary trypsinogen-2 test strip in the diagnosis of acute pancreatitis.

OBJECTIVES: Early diagnosis of acute pancreatitis remains a challenge. A rapid dipstick screening test for acute pancreatitis has been developed. This prospective study was designed to evaluate the diagnostic value and time course of the rapid urinary trypsinogen-2 test strip in acute pancreatitis, with comparisons with serum amylase and serum lipase. METHODS: A total of 165 patients with acute abdominal pain (67 with acute pancreatitis and 98 with other acute abdominal diseases) attending our emergency unit were included. All patients were tested with the urinary trypsinogen-2 test strip, and serum amylase and serum lipase concentrations were determined simultaneously. To measure the time course of the urinary trypsinogen-2 test, 32 patients with acute pancreatitis were tested with a urinary trypsinogen-2 test strip on days 1, 2, 3, and 4 after admission. RESULTS: Using a cutoff level of 50 microg/L for urinary trypsinogen-2, the sensitivity, specificity, and accuracy of the urinary trypsinogen-2 test strip for recognition of acute pancreatitis were 89.6%, 85.7%, and 87.3%, respectively. The diagnostic accuracy rates of serum amylase and serum lipase were 88.5% and 93.3%, using cutoff values of 3 times the upper normal limits for serum amylase and serum lipase, respectively. All but one of the 17 patients with severe acute pancreatitis was detected by the test strip (sensitivity, 94.1%). The time-course study of the urinary trypsinogen-2 test strip revealed that the sensitivity on days 1, 2, 3, and 4 was 90.6%, 81.2%, 59.4%, and 50%, respectively. There was no significant difference in the sensitivity between urinary trypsinogen-2 and serum lipase; however, the sensitivity values of serum lipase were significantly higher than those of serum amylase from days 1 to 4. CONCLUSION: The rapid urinary trypsinogen-2 test is a reliable and simple method for the early diagnosis of acute pancreatitis. A positive test identifies patients in need of further diagnostic measures. The urinary trypsinogen-2 test can be performed in health care units where laboratory testing facilities are not immediately available.

Abdominal Pain↗

Modification of the tear function index and its use in the diagnosis of Sjögren's syndrome.

BACKGROUND: The tear function index (TFI) has been shown to be of value in the diagnosis of patients suffering from Sjögren's syndrome. It is dependent, however, on introducing into the conjunctival fornix the correct concentration of fluorescein in at least one and a half times the normal tear volume. The stimulus and effect of this added volume on the tear dynamics is likely to vary between individuals. These factors, together with the method of performing the test, limit its general applicability. AIM: To devise a method of performing the TFI with less variability and more general applicability. To present a theoretical and in vitro assessment of the dynamics of the TFI. METHOD: The study was divided into three parts. The first part was to compare the results obtained using a prepared strip containing 1.3 microl of 0.5% fluorescein with the introduction of the same amount of fluorescein as a drop. The second part was to compare the results obtained with prepared strips with the standard method of performing the TFI, both with and without topical anaesthetic. The third part was an in vitro study of the rate of flow of graded volumes on a filter paper strip. 42 subjects with a diagnosis of Sjögren's syndrome according to the European criteria and 126 without Sjögren's syndrome were included. RESULTS: There was no significant difference between the results obtained with a prepared strip and the introduction of 1.3 microl into the eye before performing the Schirmer's test and TFI (0.1<p<0.93). There was, likewise, no significant difference between using the prepared strips and the standard method of performing the TFI (0.36<p<0.93). There was, however, less interocular difference (p=0.01) and variability (p=0.001) using the prepared strips than using a drop of fluorescein. Patients with Sjögren's syndrome had mean TFIs of 11.7 and 8.61 with upper 95% confidence values of 15 and 12 without and with topical anaesthetic, respectively. The theoretical calculation of the TFI was similar to the observed values. The in vitro results allow the filter paper to be removed from the eye at any interval and to estimate the volume of tears that the filter paper was in contact with. CONCLUSION: The proposed method of performing the TFI is easy to perform, reliable, and therefore has general applicability for primary care and general practitioners. It allows the rapid identification of subjects who may be suffering from Sjögren's syndrome.

Aged↗

Microalbuminuria measurements by two in-house ELISA methods.

Detection of microalbuminuria is important in the management of diabetic patients since it is predictive of development of proteinuria and nephropathy. Two sensitive and specific in-house ELISAs for microalbuminuria were established and validated. One of the ELISAs was based on antigen coating while the other employed antibody coating. Recovery and linearity experiments gave acceptable results of 100 +/- 10%, while precision results were <10% for intra-assay and <12% for inter-assay coefficients of variation (CVs). The standard curve ranged from 10-625 ug/l, equivalent to 0.2-12.5 mg/l for urine samples diluted 1:20 fold. When the antibody coated ELISA was compared to antigen coated ELISA, a correlation of r=0.996 was obtained. When compared to commercial kits, the in-house ELISAs gave good correlations of r=0.961 versus the Boehringer Mannheim Micral Test strips and r=0.940 versus Ames Microalb Turbidimetry. The normal microalbumin reference ranges determined for 12h, first morning and random urine samples were 0.7-5.3 mg, 0.1-10.2 mg/l and 0.8-26.1 mg/l respectively. The normal albumin excretion rate (AER) was 1.0-7.3 ug/min while untimed urine samples gave results of 0.1-0.9 and 0.2-1.6 mg/mmol after dividing by creatinine concentrations. The ELISAs were used to detect microalbuminuria in 338 random urine samples from diabetic patients. A high percentage 47.9% was found to be positive for microalbuminuria and 18.0% had macroalbuminuria >25 mg/mmol. Thus screening for microalbuminuria together with creatinine measurements using random urine samples can be used for management of diabetic patients.

Adolescent↗

Evaluation of Xenostrip-Tv, a rapid diagnostic test for Trichomonas vaginalis infection.

An immunochromatographic strip test, Xenostrip-Tv, was compared to wet mount and PCR for the diagnosis of Trichomonas vaginalis infection in women. Of 428 specimens tested, 54 (12.6%) were positive by an "expanded gold standard," defined as either a positive wet mount and PCR test with primers TVK3 and TVK7 and/or a positive PCR test confirmed by a second PCR assay with primers TVA5-1 and TVA6; 26 (6%) were positive by wet mount, and 36 (8.4%) were positive by Xenostrip-Tv test. Since the Xenostrip-Tv test is rapid and easy to perform and proved to be more sensitive than wet mount, it should be considered as an alternative to wet mount for point-of-care diagnosis of trichomoniasis, especially in settings where microscopy is impractical.

Animals↗

New approach to technical and clinical evaluation of devices for self-monitoring of blood glucose.

Despite the wide distribution of devices for self-monitoring of blood glucose (SMBG), there is no internationally accepted agreement on a standardized procedure for their evaluation. This is due to incomplete or even inappropriate technical evaluation and to inadequate evaluation criteria for their clinical acceptability. To provide adequate information on the performance of these devices over the whole clinically relevant range for SMBG (30-350 mg/dl), a standardized test procedure has been established for technical (accuracy, precision, and total deviation) and clinical (acceptance analysis) evaluation. To demonstrate the potency of this new approach, the following SMBG devices and test strips were evaluated: Chemstrip bG batch (n = 10), Glucostix batch (n = 2), Accu-Check II (n = 5), Diascan (n = 5), Diascan strip batch (n = 4), Glucocheck SC (n = 4), and Glucometer II (n = 4). The devices and test strips were examined by trained technicians, and in addition, 1 Chemstrip bG batch and 2 Accu-Check II meters were examined by 10 and 5 trained diabetic patients, respectively. Even the best-performing device did not achieve the American Diabetes Association's goal that SMBG measurements should be within 15% of the reference value. Instead, the maximal total deviation within the clinically relevant blood glucose range reached values that equalled deviations from the reference value between 16 and 108%. Three of 36 devices were classified as good, 29 as acceptable, and 4 as unacceptable for clinical use. In conclusion, this new approach to the technical and clinical evaluation of devices for SMBG is easy to perform and provides more realistic and comparable information for clinical use and approval than commonly used methods.

Blood Glucose Self-Monitoring↗

Use of rapid dipstick tests to exclude urinary tract infection in children.

Children presenting with symptoms attributable to urinary tract infection (UTI) are not uncommonly referred to paediatric departments for assessment. The aim of this study was to evaluate the use of rapid dipstick tests in the diagnosis of urinary tract infection in children. Urine was collected from 375 children admitted to a general paediatric ward, in whom UTI was a possibility on clinical grounds. Of these, 124 were less than one year old. Urine was tested with a dipstick for the presence of nitrite and leucocyte esterase. Bacterial culture and examination for white cells, red cells and other formed elements were performed. The results of the dipstick tests, microscopy and culture were correlated with the clinical details. Combination of a negative dipstick test for nitrite and leucocyte esterase showed a negative predictive value for UTI of 96.9% and a specificity of 98.7%. In children less than a year old these values were 96.7% and 99.2% respectively. The leucocyte esterase strip test showed a negative predictive value for pyuria of 94.3% with a specificity of 86.9%. In children less than a year old these values were 93.1% and 84.4% respectively. The use of dipsticks for the detection of urinary nitrate and leucocyte esterase in daily clinical practice is recommended. In children, the absence of both nitrite and leucocyte esterase in urine indicates that UTI is unlikely; however, positive dipstick tests for nitrite and/or leucocyte esterase are not specific indicators of UTI, and should not be used in place of laboratory examination. The dipstick method is most likely to be useful as a screening test to exclude UTI in children, but may be less suitable for infants. It should not be used to diagnose urinary tract infection.

Adolescent↗

Effect of mixed mutans streptococci colonization on caries development.

OBJECTIVE: To evaluate the clinical importance of mixed mutans streptococci colonization in predicting caries in preschool children. METHODS: Caries prevalence was examined twice, with a 6-month interval, in 410 preschool children aged 3-4 years at baseline. A commercial strip method was used to evaluate the mutans streptococci score in plaque collected from eight selected interdental spaces and in saliva. Mutans streptococci typing polymerase chain reaction (PCR) assays (Streptococcus sobrinus and Streptococcus mutans, including serotypes c, e, and f) were performed using colonies on the strips as template. RESULTS: Twenty variables were examined in a univariate analysis to predict caries development: questionnaire variables, results of clinical examination, mutans streptococci scores, and PCR detection of S. sobrinus and S. mutans (including serotypes c, e, and f). Sixteen variables showed statistically significant associations (P < 0.04) in the univariate analysis. However, when entered into a logistic regression, only five variables remained significant (P < 0.05): caries experience at baseline; mixed colonization of S. sobrinus and S. mutans including S. mutans serotypes; high plaque mutans streptococci score; habitual use of sweet drinks; and nonuse of fluoride toothpaste. CONCLUSION: 'Mixed mutans streptococci colonization' is a novel measure correlated with caries development in their primary dentition.

Beverages↗