• Journal of Veterinary Clinics
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• v.26 no.2
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• pp.130-133
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• 2009

The use of screening tests as part of a diagnostic work-up is common in domestic canine practice, but understanding of the diagnostic test characteristics and factors affecting diagnostic accuracy is not clear among clinicians. This article was aimed to provide clinicians with a better understanding on the selection of test kits and with a proper interpretation of test results using an example from heartworm(Dirofilaria immitis) studies. From the literatures, diagnostic accuracy varied depending on the kits: percent average sensitivity and specificity of ELISA antigen-detecting kits were DiroChek(Synbiotics, USA) 78.1 and 95.2, SNAP(IDEXX, USA) 66.3 and 98.1, and Solo Step(Heska, Switzerland) 69.5 and 97.5, respectively, while the values for three hematological methods(Modified Knott's, direct smear and capillary tube) ranged from 38.4 to 81.8% and from 96.9 to 100%, respectively. Furthermore, it was also reported that the prevalence of heartworm disease in domestic dog populations varied depending on the regions studied: 2.5-22.8% for microfilarial test and 2.2-66.3% by ELISA. The values of predictive values for positive(PPV) and negative(NPV) provide useful information to clinicians on the probability of heartworm infection, but the PPV and NPV are greatly dependent on the heartworm prevalence. This suggests that PPV or NPV values of a test should be interpreted carefully in different clinical settings. Practical methods on the interpretation taking into account heartworm prevalence were discussed.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.77-81
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• 2007

Diagnostic performance of polymerase chain reaction (PCR) for detecting Dirofilaria immitis in dogs was evaluated when no gold standard test was employed. An enzyme-linked immunosorbent assay test kit (SnapTM, IDEXX, USA) with unknown parameters was also employed. The sensitivity and specificity of the PCR from two-population model were estimated by using both maximum likelihood using expectation-maximization (EM) algorithm and Bayesian method, assuming conditional independence between the two tests. A total of 266 samples, 133 samples in each trial, were randomly retrieved from the heartworm database records during the year 2002-2004 in a university animal hospital. These data originated from the test results of military dogs which were brought for routine medical check-up or testing for heartworm infection. When combined 2 trials, sensitivity and specificity of the PCR was 96.4-96.7% and 97.6-98.8% in EM and 94.4-94.8% and 97.1-98% in Bayesian. There were no statistical differences between estimates. This finding indicates that the PCR assay could be useful screening tool for detecting heartworm antigen in dogs. This study was provided further evidences that Bayesian approach is an alternative approach to draw better inference about the performance of a new diagnostic test in case when either gold test is not available.

• The Journal of the Korean Society for Microbiology
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• v.15 no.1
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• pp.9-17
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• 1980

In the ecology and epidemiologic studies on various serotypes of atypical mycobacteria(AM), Schaefer's bacterial agglutination test(BA) provided the basis of the serologic procedures. Recently, attempts have been made to modify and to simplify the Schaefer's BA such as a slide agglutination test(Engel & Beerwald, 1970), a "simplified" BA(Reznikov & Leggo, 1972), an agglutination inhibition test(Richards & Eacret, 1972) and "micromethod"(Thoen et al., 1975). The BA, however, was not widely applied as a routine laboratory test mainly because it requires much times and labors to perform and partley because it is not applicable to hydrophobic strains either often encountered in the isolation of AM in the clinical bacteriology or stock strains maintained in the laboratory. On the contrary, fluorescent antibody technique with mycobacteria may have advantages over the BA because it is far more simpler in serologic procedures and is applicable to all strains of mycobacteria regardless of smooth or rough types of cultures. At the present, it is well known that the type-specific antigens are lacking on the surface of rough type of AM compared to that on smooth type of strain, but the antigenicity on the surface of the hydrophobic strains of AM which resulted from a series of subculture and the strain in the laboratory for 3 to 6 months has not been clarified. In this study, an attempt to serotype the hydrophobic strains of M. scrofulaceum serotype 41, 42 and 43 by fluorescent anti-complement(FAC) technique was made. The FAC technique with mycobacteria was also described in detail. In the summary, the complement fixing antibody titres of reference sera to smooth types of homologous serotype was highest, but the antibody titres of reference sera to hydrophobic strains of serotypes, 41, 42 and 43 gave two-to 8-folds lower than those to smooth type of strains. Although the sensitivity of type-specific antigens on the hydrophobic strains to reference sera was much lower, using the two units of reference sera determined by titration with hydrophobic strains, three serotypes, i. e., 41, 42 and 43 were specifically differentiated one another by FAC technique. This result indicated that the hydrophobic strains which were maintained in the laboratory at least for 6 months still retain type-specific antigen detectable by FAC technique.

• Journal of Veterinary Clinics
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• v.26 no.4
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• pp.311-316
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• 2009

The purpose of this meta-analysis was to collect data from the literatures reporting prevalence of canine heartworm (HW) infection in an effort to emphasize the preventive importance of the disease and to serve as baseline data for further studies related to the disease management. Fourteen studies regarding the prevalence of HW infection published during the 1996-2007 were identified using the meta-analytic approach, involving a total of 1,790 cases by microfilarial test and 3,277 cases by enzyme-linked immunosorbent assay (ELISA) antigen test for adult worms. Data analysis was performed separately for pet and military dogs. The overall apparent prevalence of HW infection in pet dogs by microfilarial test and ELISA was 11.7% (183/1,563) and 16.5% (379/2,302), respectively (p < 0.001). No significant difference in prevalence by microfilaria and ELISA was observed between sexes whether assessed by data separately or pooled together. Compared to before 2000, the prevalence by microfilaria after 2000 has increased from 2.5% to 13.1% (p < 0.001), and the rate of adult worm has also increased from 9.9% to 20.3% (p < 0.001). The prevalence determined by ELISA was found to increase with age: 20.7% (n = 1,003) of dogs older than 2 years were positive, compared with 5.0% of dogs (n = 701) under 2 years of age (p < 0.001). Also, outdoor dogs showed significantly higher prevalence than indoor dogs (37.9% versus 3.4%; p < 0.001). For military dogs, the overall prevalence was 38.3% (87/227; 95% CI, 32.1 - 44.7%) by microfilarial test and 38.5% (375/975; 95% CI, 35.4 - 41.6%) by ELISA. This rate was significantly higher than that of pet dogs (p < 0.001). Occult infection rate of 37.5% and 66.7% was found in pet and military dogs, respectively.

• The Korean Journal of Urological Oncology
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• 제15권3호
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• pp.131-136
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• 2017

Purpose: The aim of this study is to investigate the criteria of biochemical recurrence (BCR) and follow-up periods and methods with and without blood and imaging test of urologic oncology before established guidelines of prostate cancer in Korea. Materials and Methods: In December 2015, we sent the questionnaire to urologic oncologist in academic hospital and received the answer from 108 urologic oncologist (50%). Also, we analyzed the data of 1,141 patients underwent radical prostatectomy in 2005 from Korean Medical Insurance. Results: In follow-up, 72 physicians (66.7%) performed blood test every 3 months, 51 physicians (47.2%) performed imaging study in case of BCR. Bone scan was the most common imaging study in the follow-up (74 physicians, 68.5%). But, bone scan was only performed in case of BCR (43 physicians, 39.8%). The criteria of BCR was PSA 0.2 ng/mL (75 physician, 69.4%), 76 physicians (70.4%) was performed different follow-up according to risk of patients. In Korean Medical Insurance data analysis, PSA were performed average 2 times every year and magnetic resonance imaging, computed tomography, Bone scan were performed average 0.1, 0.2, 0.1 times every year, respectively. Conclusions: The criteria of BCR and the follow-up of prostate cancer patients in Korea were similar Korean prostate cancer guidelines. Blood and imaging test might be increased compared to 10 years ago, it is necessary to compare the Korean Medical Insurance data between 10 years ago and present.

• Parasites, Hosts and Diseases
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• v.24 no.1
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• pp.25-41
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• 1986

The applicability of micro-ELISA was evaluated in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of $2.5{\;}{\mu}g/ml$. Serum was diluted to 1 : 100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of o. 18 in both samples. The results were summarized as follows: 1. The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1% ; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. 2. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5%; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. 3. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 ot 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. 4. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. 5. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes.

• Journal of fish pathology
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• v.6 no.1
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• pp.11-20
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• 1993

To study the immune responses of the japanese eel. Anguilla japonica, fish were injected intraperitoneally with several types of Edwardsiella tarda antigen, i. e., FKC(formalin killed cells), HKC(heat killed cells) or LPS(lipopolysaccharide), and the changes of immunocytes numbers, phagocytosis and agglutination titre in the peripheral blood of the fish were investigated. The number of lymphocytes in the peripheral blood of eels were decreased until 6 hours after injection, and then were turn to normal levels after 24 hours of injection. However, the level were slightly increased and were remained after 24 hours. The number of neutrophils of FKC, HKC or LPS injected fish were the highest at 12 hours after injection and were decreased slowly after that. Three weeks after the injections, the agglutination of antibody titre of all immunized groups were reached at 128 and were remained this level thereafter. However 6 weeks after the injections, that in HKC injected fish were dropped the level up to 4. Fish were injected with LPS and the blood from the fish were bled after 12 hours. Then the blood were incubated with E. tarda. Six hours after incubation, the phagocytic index was reached the highest level, 28.3. One week after the LPS injection, the blood were again bled and incubated with E. tarda. The phagocytic index at this time was 3.9. The phagocytic indexes of the fish injected with FKC and HKC, treted as same LPS injected fish as above, were 18.8 and 10.7, respectively. The phagocytic index of the control fish was 1.2. The antibacterial activities of normal antiserum against E. tarda were shown for both FKC and LPS injected fish, but not for HKC injected fish. The RPS(relative percentage of survival) of HKC, FKC and LPS injected fish in the challenge test were 10%, 20% and 30%, respectively. These results suggest that the effect of protection of the eel which were injected with antigen were varied with the method of preparation of the antigen.

• Parasites, Hosts and Diseases
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• v.29 no.4
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• pp.339-354
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• 1991

Localization and characterization of the antigenic components of sparganum which induced IgG and IsM antibodies in the host were studied by immunohistochemical techniques and SDS-PAGT and Western blotting. The antigen recognized by IgG antibody of rats or mice which were immunised by infection or injection of crude extracts of metacestodes of Spirometra erinacei, was located in the parenchyme of sparganum, especially at the cortex and around the calcareous corpuscles. The immunoreaction was demonstrated not only in the encysted fibrous wall of host but around the arterioles or venules in the connective tissue of host. The antigen recognized by IgM antibody of rats or mice was also observed in the parenchyme of sparganum and in the connective tissue of host. By 5∼20% gradient SDS-PAGE and EIBT, we detected antigenic components by IgG and 1gG antibodies of the rat or mouse immunized by infection or injection of crude extract of spargana. Twenty-three antigenic bands from crude extracts of spargana were recognized by IgG antibody and 15 components by IgM antibody of immunized rats. Out of the bands recognized by IgG and IgM antibodies, 15 were cross-reacted each other. Twenty components of eBlcretory-secretory proteins from spargana were recognized by IgG, and 5 components by IgM antibody of immunized rats. By IgG and IgM antibodies of immunized mice, 16 components of crude extracts were recognized by IgG antibody and 9 components by IgM antibody. Twenty components of excretory-secretory preparation were recognized by IgG antibody and 5 components by IgM antibody. Thirteen components of crude extracts were cross-reacted by IgG antibody of rats and mice.

• Journal of Veterinary Clinics
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• v.29 no.4
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• pp.309-314
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• 2012

The study was undertaken to evaluate sensitivity and specificity of rapid Avian Influenza (AI) and Newcastle Disease virus (NDV) combo antigen kits from field samples of domestic (broiler and layer chicken, native chicken) and semi-domestic (duck, goose, pigeon and quail) birds of Bangladesh. Samples were collected from naturally infected AI suspected domestic and semi-domestic birds of five different outbreak areas in Bangladesh. From each area two birds were selected for sampling, and from each bird three types of samples (tracheal, cloacal and oro-nasal swabs) were collected. A total of 210 field samples from a total of 70 birds were collected and tested using AI and NDV combo antigen rapid diagnostic kits in the study. All three different samples from a bird showed similar pattern of reaction. Out of 210 samples, 15 samples (5 birds), 63 samples (21 birds) and 27 samples (9 birds) were positive for AIV, NDV and both for AIV and NDV, respectively; whereas the remaining birds were negative for either AIV or NDV in this screening test. Among the five AIV positive, a layer chicken from wet market in Mymensingh, Netrokona, Gibandha and Kurigram and a native chicken from wet market in Kurigram area was positive to AIV. The semi-domestic birds are either positive to NDV or free from both AIV and NDV. This study revealed that the AIV and NDV rapid diagnostic kits could be effectively use to diagnose the respective virus in trachea, oro-nasal and cloacal samples simultaneously. AIV-NDV combo Ag test result clearly indicates that the test kit designed for AIV and NDV could diagnose the disease rapidly with less effort and higher scientific know how which could be used for the detection of AIV and NDV using field samples in large scale.

• Laboratory Medicine Online
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• v.1 no.1
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• pp.26-34
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• 2011

Background: Laboratory diagnosis of new influenza A (H1N1) is crucial for managing patients and establishing control and prevention measures. We compared the diagnostic accuracies of the real time RT-PCR (rRT-PCR) test recommended for the confirmation of the new flu and the viral culture method used conventionally for viral disease with that of the rapid antigen test (RAT). Methods: We performed RAT, R-mix culture, and real-time PCR by using 861 respiratory samples collected from December 2009 to January 2010 and evaluated the abilities of these methods to detect new influenza A. The relationship among the positive rates of RAT, grades of culture, and the cycle threshold (Ct) values of rRT-PCR was also evaluated. Results: Of the 861 patients, 308 (35.8%) were diagnosed with new influenza A. The sensitivities, specificities, positive predictive values, and negative predictive values of the tests were respectively as follows: 59.7%, 99.5%, 98.4%, and 81.6% for RAT; 93.2%, 100%, 100%, and 96.3% for R-mix culture; and 95.8%, 100%, 100%, and 97.7% for rRT-PCR. Samples with weak positive grade in culture and those with Ct values of 30-37 in rRT-PCR showed positivities as low as 25.3% and 2.3% in RAT, respectively. The hospitalization rate and death rate of the confirmed patients were 3.2% and 0.3%, respectively, and gastrointestinal symptoms were observed in 7.2% of the patients. Conclusions: R-mix culture and rRT-PCR tests showed excellent reliability in the diagnosis of new influenza A and could be very useful, especially for samples with low viral load.