Comparison of the detection of Helicobacter pylori infection by commercially available serological testing kits and the 13C-urea breath test.

BACKGROUND: Serum Helicobacter pylori (H. pylori) antibody kits (LZ and LIA) using the latex agglutination immunoassay method are commercially available, but few studies have been performed to determine their diagnostic accuracy or to compare their results with those of enzyme-linked immunosorbent assay (ELISA) kits (EP and EIA). METHODS: Sera were obtained from 213 hospital outpatients with dyspeptic symptoms. The serological results were compared with the result of the 13C-urea breath test (UBT) which seems to be reliable. RESULTS: Of the 213 subjects, 154 were diagnosed as positive for H. pylori infection according to the UBT. The sensitivities and specificities of these tests were 97.4% and 76.3%, 98.1% and 78.0%, 99.4% and 74.6%, and 98.1% and 71.2% for the EP, LZ, EIA and LIA tests, respectively. When the 13 subjects whose seropositive results of the four kits were completely opposite to the negative results of the UBT were excluded, the specificities of evaluated kits were all higher than 90%. The concordance rate between the EP and EIA tests was 98.1% (Spearman's rank correlation coefficient = 0.83) and that between the LZ and LIA tests was 97.1% (correlation coefficient = 0.91). The LZ gave higher antibody titer value than EP (p < 0.0001, Z = 9.82; Wilcoxon signed-rank test), and EIA gave higher value than LIA (p < 0.0001, Z = 6.43; Wilcoxon signed-rank test). CONCLUSIONS: The latex immunoassay method provided the same reliability to ELISA in terms of the diagnostic accuracy for current H. pylori infection, although we should take into account the titer value differences by each test method in practical use.

Impact of technical training on rapid antigen detection tests (RADT) in group A streptococcal tonsillopharyngitis.

Rapid antigen detection tests (RADT) are widely used for the rapid diagnosis of group A streptococcal (GAS) tonsillopharyngitis. In a prospective 3-year study, the reliability of two different RADT methods was compared, as performed by lab technicians versus physicians. Sensitivity and specificity, as well as positive and negative predictive values, were calculated. When performed by physicians, the results (44.4 %, 8.3 %, 26.7 % and 16.7 %) of a latex agglutination test (LAT) were unacceptably low. However, after switching to a lateral-flow immunoassay (LFIT) and implementing additional hands-on training, the performance improved dramatically (100 %, 92.6 %, 84.6 % and 100 %). In conclusion, technical errors, along with a lack of experience and expertise, negatively impact RADT accuracy.

Evaluation of conventional & serological methods for rapid diagnosis of cryptococcosis.

BACKGROUND & OBJECTIVE: Cryptococcosis is a chronic infective condition affecting the central nervous system. Unless diagnosed early and specific treatment instituted it can be fatal. There is an urgent need for a rapid and specific diagnostic tool for better management of the patients. Conventional methods such as culture and India ink are specific but cumbersome and time consuming. Serological methods of detection are rapid but have problems of false positivity and cross-reactivity with other micro-organisms. We carried out this study to compare and evaluate the conventional methods with serological methods of detection of cryptococcal meningitis. METHODS: A comparative evaluation of conventional methods (India ink and culture) with LAT (latex agglutination test) and EIA (enzyme immunoassay) was done in 127 CSF samples using culture and EIA as reference separately. RESULTS: India ink was positive for Cryptococcus in 72.4 per cent of the samples; 56 per cent were culture positive; LAT positive were 85 per cent and 79.5 per cent were positive by EIA. When culture was positive, all other tests were in agreement to it. However, when culture was negative there was significant difference between the pair of discordance of various diagnostic tests. Culture was 83.46 per cent in agreement to India ink, 76.3 per cent to EIA and 70.8 per cent to LAT. EIA was 92.9 per cent in agreement to India ink and LAT; 6.3 per cent showed false positive by LAT. INTERPRETATION & CONCLUSION: EIA is valuable in establishing diagnosis when culture is negative for cryptococcosis. EIA is more specific and has potential advantages over LAT as it gives clear discrimination of positive from negative results. Thus, EIA may be used as a simple, rapid, and reliable serological test for early detection of cryptococcal antigen in clinical samples like CSF in routine laboratories.

A comparative study of optical techniques applied to particle-enhanced assays of C-reactive protein.

This work is based on the well-established immunoassay principle of agglutination of latex particles covered by immunoproteins. In our experiments, positively charged particles act as carriers for the F(ab')2 fragment, obtained from rabbit polyclonal IgG, active against C-reactive protein (CRP). The presence of the antigen CRP in the immunolatex system causes agglutination and the aim of the present study was to compare different optical techniques (turbidimetry, nephelometry, angular anisotropy and photon correlation spectroscopy) capable of detecting the agglutination. The sensitivity and detection limit largely depend on the optical method. We have analyzed for each optical technique the following aspects: sensitivity, reproducibility, detection limit, reaction time, amount of sample wasted and availability of the required detection device. The results presented in this paper show that both angular anisotropy and photon correlation spectroscopy offer lower detection limits, and use little reagent, but have longer assay times than the classical optical techniques of turbidimetry and nephelometry.

More laboratory testing: greater cost but not necessarily better.

BACKGROUND: The bacterial latex agglutination assay is ordered predominantly on the pediatric population, for rapid screening for bacterial surface antigens in cerebrospinal fluid (CSF) or urine specimens. The high cost of this assay and questions raised in the literature regarding its accuracy led to a retrospective review of the use of this assay at a medium-sized midwest teaching hospital. The results of 6,370 bacterial latex agglutination tests performed between May, 1995, and November, 1996, and charts of patients being tested were reviewed. RESULTS: This study demonstrated a sensitivity and specificity of 28.6% and 86.7% for urine specimens and 70.0% and 99.4% for CSF specimens. A total of 11 pathogens were accurately detected (7 CSF and 4 urine). There were 13 false negatives and 59 false positives. None of the true positives had a discernible effect on either treatment or hospital course; however, several of the erroneous tests resulted in delayed or unnecessary treatment and workup of the involved patients. The annual billed cost of this test at this institution (fiscal years 1995 to 1997) averaged $167,000 per annum. This does not include indirect costs associated with increased length of hospital stay, overutilization of antibiotics and excess laboratory tests ordered as a result of false positives. CONCLUSIONS: Bacterial antigen latex agglutination testing is neither sufficiently sensitive nor specific to be used as a screening test. Accurate results have no demonstrable clinical impact, whereas numerous inaccurate results are often generated at great cost. The continued use of the latex agglutination assay should be seriously questioned in an era when cost containment and clinical efficiency are becoming increasingly important.

Comparison of an enzyme immunoassay and a latex agglutination system for the diagnosis of invasive aspergillosis in bone marrow transplant recipients.

The performance of two Aspergillus antigenemia systems, the sandwich enzyme-linked immunosorbent assay (ELISA), Platelia Aspergillus test, and the latex agglutination (LA), Pastorex Aspergillus test, in the diagnosis of invasive aspergillosis were compared by testing 364 serum samples from 22 bone marrow transplant (BMT) recipients. Sensitivity and specificity for the ELISA test were 60% and 82% respectively, vs 40% and 94% for the LA test. In the two patients found positive with both methods, the ELISA test became positive earlier than the LA test or remained positive after the LA test had become negative. These results encourage further evaluation of the Platelia Aspergillus test, to assess its role in the management of invasive aspergillosis in BMT patients.

Detection of viruses in environmental samples: suitability of commercial rotavirus and adenovirus test kits.

Commercially marketed kits are now available for rapid viral assay of clinical specimens. This study was conducted to determine the suitability of these kits for use in environmental testing. Eight rotavirus kits and one enteric adenovirus kit were screened for sensitivity using simian rotavirus SA11, human rotavirus Wa, and adenovirus 41. The most sensitive rotavirus kit and the adenovirus kit were selected for further evaluation using virus-seeded and unseeded sewage samples. The selected rotavirus kit proved capable of detecting virus at the 10(1) PFU/ml level. The enteric adenovirus kit was similarly sensitive, detecting virus at the 10(1) TCID50/ml level. Neither kit was adversely affected by the presence of sewage. Kit assay revealed 3 of 30 unseeded sewage samples to be positive for rotavirus. Adenovirus positive samples were not detected among the 30 samples. These results were confirmed using electron microscopy. It was concluded that sensitive commercial kits could provide a reasonable alternative to cell culture for the presumptive testing of environmental samples.

Clinical evaluation of the BTA TRAK assay and comparison to voided urine cytology and the Bard BTA test in patients with recurrent bladder tumors. The Multi Center Study Group.

OBJECTIVES: To assess the clinical performance of the BTA TRAK assay and to compare it with that of voided urine cytology (VUC) and the Bard BTA test (BTA) in the detection of recurrent bladder cancer (BC). METHODS: The study was performed on randomly selected archival voided urine samples for many of which VUC and/or BTA information was available. Sensitivity was determined in samples from patients with histologically confirmed recurrent BC. Specificity was determined in samples from healthy volunteers, patients with three categories of current medical conditions, and patients with a history of BC but no current evidence of disease. RESULTS: The TRAK assay was positive in 156 of 216 samples for patients diagnosed with BC, for an overall sensitivity of 72%. Mean values increased with progressing grade and stage of disease. In the comparison between TRAK and VUC, the overall sensitivities were 68% and 25%, respectively (P < 0.001). For Stages Ta and T1 and for all tumor grades, the sensitivity of the TRAK assay was significantly greater than that of VUC (P < 0.001). TRAK sensitivity was also significantly better than that of BTA (73% versus 58%, P = 0.005). The specificity of the TRAK assay ranged from 75% in samples from patients with genitourinary disease to 97% in healthy volunteers. CONCLUSIONS: The TRAK assay is superior to VUC and the original BTA test in the detection of BC. The results of the study indicate that the TRAK assay may be a useful adjunct to cystoscopy in the management of patients with recurrent BC.

Sensitivity of intrapartum group B streptococcal screening and in vitro comparison of four rapid antigen tests.

OBJECTIVES: To evaluate the sensitivity of intrapartum screening for group B streptococcal (GBS) colonization and to compare 4 rapid GBS antigen tests in vitro. DESIGN: Two swabs of the lower vagina of 769 parturients were taken; one swab was cultured, the other was frozen at -70 degrees C until antigen testing with the Group B Strep Test (Quidel) of the culture positive samples was performed. The Quidel test was then compared with 3 other rapid GBS antigen tests in vitro: Wellcogen Strep B (Wellcome Diagnostics), Slidex méningite Strepto B (bioMérieux) and ICON Strep B (Hybritech). The supernatant of 29 GBS cultures in Todd-Hewitt broth was tested in bacterial concentrations of 10(6), 10(7), and 10(8) Colony-forming Units (CFU)/ml, respectively. RESULTS: Lower vagina GBS carrier rate was 13.4% (103/769) and heavy colonization (growth density 3 and 4 on blood agar plates) was found in 5.2% (40/769). The Group B Strep Test detected 11% (11/103) of GBS carriers, with a sensitivity for heavy colonization of 25% (10/40). In vitro none of the tests scored positively with a concentration of 10(6) CFU/ml, while with 10(7) CFU/ml the enzyme immunoassay tests (Quidel, Hybritech) were more sensitive (McNemar test, P < 0.05) than the latex agglutination tests (Wellcome Diagnostics, bioMérieux). CONCLUSIONS: Although in vitro the enzyme immunoassay tests are more sensitive than the latex agglutination tests, sensitivity in vivo is too low to recommend the use of rapid antigen tests for general screening.

Chlamydia latex agglutination antigen and protocol improvement and psittacine bird anti-chlamydial immunoglobulin reactivity.

Latex agglutination detection of chlamydial antibody activity in psittacine bird sera was significantly more sensitive when an improved protocol was followed than was a test using the previously used protocol. Titers of antibody-positive sera were fourfold to 32-fold higher by the improved protocol than titers by the previously used protocol, whereas antibody-negative sera were negative by both protocols. Column chromatography was used to separate immunoglobulins in psittacine bird serum. Immunoglobulin M was reactive in latex agglutination (LA) but non-reactive in direct complement fixation (DCF). Immunoglobulin G was reactive in LA and DCF. The fifth supernatant fluid of serum multiply absorbed with latex beads was non-reactive in LA and DCF. The immunoglobulins reactive in LA had high avidity and affinity for latex beads. Prolonged storage at 4 C to 6 C preserved LA reactivity of absorbed immunoglobulins better than storage at 21 C to 23 C.

[The diagnostic evaluation of rapid sputum technics for Pneumococcus in community-acquired pneumonia. The usefulness of Bayes theorem for clinical application]. / Evaluación diagnóstica de las técnicas rápidas del esputo frente neumococo en la neumonía adquirida en la comunidad. Utilización del teorema de Bayes para su aplicación clínica.

This study aimed to quantify the diagnostic value of immunological techniques and methods for rapid analysis of sputum for pneumococcus, using sensitivity and specificity values reported in the literature to calculate positive and negative predictive values (PPV and NPV) according to Bayes formulas. Diagnostic gains of the test are calculated and compared to pretext probability. We located articles reporting sensitivity and specificity of counterimmunoelectrophoresis (CIE), coagglutination (CoA) and latex agglutination (LA) tests. We also calculated the probability ratios for the three tests. LA achieved the best overall diagnostic utility rating. CoA had the highest PPV, whereas LA offered the highest NPV. CIE was the least useful. These three tests are more useful at intermediate levels of prevalence of pneumococcus, which coincide with estimate in our population. We conclude that LA and CoA are of greater diagnostic utility for community acquired pneumonia, as they are useful for determining prevalence as well as for deciding initial antibiotic treatment.

Comparison of four latex kits for detection of E. coli O157.

OBJECTIVE: To compare four Escherichia coli O157 test kits for detection of E. coli O157:H7 isolated from clinical specimens. DESIGN: One hundred two Escherichia coli O157:H7 isolates obtained from stored specimens and 99 non-sorbitol fermenting enterobacteriaceae isolates from current clinical specimens were tested against four latex kits: Wellcolex, RIM, Prolex, and Oxoid. Each isolate was tested against all four kits on the same day. SETTING: Provincial Laboratory of Saskatchewan, Canada. PATIENTS: Patients from Saskatchewan with diarrhea submitted stool specimens through their family physicians to the Provincial Laboratory for detection of enteric pathogens including E. coli O157:H7. RESULTS: The sensitivity and specificity of each test kit were: Wellcolex 100%, 99%; RIM 100%, 99%; Prolex 99%, 100%; Oxoid 100%, 100%. The Prolex kit failed to detect one E. coli O157:H7 isolate. CONCLUSION: All kits tested were able to identify E. coli O157 isolated from stool specimens. Further study with Prolex is needed to assess the significance of the one missed E. coli O157 isolate.

[The demonstration of specific Salmonella typhi antigens by the latex agglutination method]. / Indikatsiia spetsificheskikh antigenov Salmonella typhi metodom lateksnoi aggliutinatsii.

The evaluation of the diagnostic importance of the method of latex agglutination with antibody diagnostica for the detection S.typhi O and Vi antigens in biosubstrates obtained from patients is presented. The clinical approbation of the method has made it possible to characterize its diagnostic specificity (whose indices varied from 95.8 to 100%), sensitivity and significant diagnostic effectiveness (67.7-91.3%). The method has been recommended for the early diagnosis of typhoid fever.

[The determination of class-M immunoglobulins in the latex agglutination reaction]. / Opredelenie immunoglobinov klassa M v reaktsii aggliutinatsii lateksa.

Results of measuring the concentrations of IgM in human serum using a specific latex diagnostic agent in the latex agglutination test (LAT) are presented. The authors demonstrate a higher efficacy of LAT in comparison with Mancini's method.

[The production of immunochemical reagents for the determination of C-reactive protein by latex agglutination]. / Poluchenie immunokhimicheskikh reagentov dlia opredeleniia C-reaktivnogo belka metodom lateksnoi aggliutinatsii.

A new simple and convenient method for the detection of C-reactive protein (CRP) in patients' blood sera is based on latex agglutination making use of new compounds obtained by conjugation of IgG fraction containing goat antibodies to CRP with various latex particles. The sensitivity of the methods is 3 mg/ml, this permitting its wide clinical application.

[The determination of opiates by latex agglutination]. / Opredelenie opiatov metodom lateksnoi aggliutinatsii.

A method based on latex agglutination test has been developed for detection of opiates in human urine. Specific reagents were synthesized and conditions of interaction of conjugated latex antigens with specific antibodies to morphine in the presence of an inhibitor were examined. Sensitivity of the method was 100 ng/ml. The developed latex agglutination test for detection of opiates may be used in combination with enzyme immunoassay in practical forensic medicine and narcology.

Evaluation of a commercial latex agglutination assay for serological diagnosis of leptospirosis.

Leptospirosis is a febrile zoonosis of worldwide distribution. A latex agglutination assay was evaluated in two studies, the first using a panel of well-characterized sera from patients with leptospirosis and from patients with other disease states and the second, a prospective hospital-based study, evaluating sera from 186 consecutive patients admitted to hospital with acute febrile illness. The confirmed leptospirosis serum panel included paired acute- and convalescent-phase specimens from 40 cases, of which 34 gave positive latex tests (case sensitivity, 85%; 95% confidence interval [95% CI], 70 to 94%). The other diseases represented in the panel of 112 specimens from nonleptospirosis patients included autoimmune diseases, brucellosis, dengue, melioidosis, malaria, syphilis, toxoplasmosis, viral hepatitis, and a number of other viral infections. The specificity of latex agglutination using this panel was 81% (95% CI, 73 to 87%). Among the patients with acute febrile illness, there were 25 cases of leptospirosis and 161 patients with other diagnoses. The sensitivity and specificity of latex agglutination in this group were 88% (95% CI, 72 to 97%) and 98% (95% CI, 95 to 100%), respectively. In this evaluation, the two distinct groups of specimens gave similar results for sensitivity, but specificity was different in each study. The sensitivity and specificity observed for the hospital study were similar to those obtained in evaluations of other rapid tests in the same population. The results of this study suggest that multiple evaluations of new diagnostic assays should be performed, because performance characteristics may vary in different populations.