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The Singulex Clarity C. diff toxins A/B (Clarity) assay is an automated, ultrasensitive immunoassay for the detection of Clostridioides difficile toxins in stool. In this study, the performance of the Clarity assay was compared to that of a multistep algorithm using an enzyme immunoassay (EIA) for detection of glutamate dehydrogenase (GDH) and toxins A and B arbitrated by a semiquantitative cell cytotoxicity neutralization assay (CCNA). The performance of the assay was evaluated using 211 residual deidentified stool samples tested with a GDH-and-toxin EIA (C. Diff Quik Chek Complete; Techlab), with GDH-and-toxin discordant samples tested with CCNA. The stool samples were stored at –80°C before being tested with the Clarity assay. For samples discordant between Clarity and the standard-of-care algorithm, the samples were tested with PCR (Xpert C. difficile; Cepheid), and chart review was performed. The testing algorithm resulted in 34 GDH(+)/toxin(+), 53 GDH(−)/toxin(−), and 124 GDH(+)/toxin(−) samples, of which 39 were CCNA(+) and 85 were CCNA(−). Clarity had 96.2% negative agreement with GDH(−)/toxin(−) samples, 100% positive agreement with GDH(+)/toxin(+) samples, and 95.3% agreement with GDH(+)/toxin(−)/CCNA(−) samples. The Clarity result was invalid for one sample. Clarity agreed with 61.5% of GDH(+)/toxin(−)/CCNA(+) samples, 90.0% of GDH(+)/toxin(−)/CCNA(+) (high-positive) samples, and 31.6% of GDH(+)/toxin(−)/CCNA(+) (low-positive) samples. The Singulex Clarity C. diff toxins A/B assay demonstrated high agreement with a testing algorithm utilizing a GDH-and-toxin EIA and CCNA. This novel automated assay may offer an accurate, stand-alone solution for C. difficile infection (CDI) diagnostics, and further prospective clinical studies are merited.

In the United States, laboratories frequently offer multiple different assays for testing of cerebrospinal fluid (CSF) samples to provide laboratory support for the diagnosis of central nervous system Lyme disease (CNSLD). Often included among these diagnostic tests are the same enzyme immunoassays and immunoblots that are routinely used to detect the presence of antibodies to Borrelia burgdorferi in serum. However, performing these assays on CSF alone may yield positive results simply from passive diffusion of serum antibodies into the CSF. In addition, such tests are only U.S. Food and Drug Administration cleared and well validated for testing serum, not CSF. When performed using CSF, positive results from these assays do not establish the presence of intrathecal antibody production to B. burgdorferi and therefore should not be offered. The preferred test to detect intrathecal production of antibodies to B. burgdorferi is the antibody index assay, which corrects for passive diffusion of serum antibodies into CSF and requires testing of paired serum and CSF collected at approximately the same time. However, this assay also has limitations and should only be used to establish a diagnosis of CNSLD in conjunction with patient exposure history, clinical presentation, and other laboratory findings.

Coccidioidomycosis (CM), a serious life-threatening fungal infection endemic to arid regions of the western United States and Mexico, can be challenging to diagnose in a timely manner. Commercially developed enzyme immunoassays (EIAs) (from Meridian Biosciences and Immuno-Mycologics [IMMY]) have provided faster, simpler means for serodiagnosis; however, independent evaluations have questioned EIA specificity, particularly IgM-positive/IgG-negative results. This study was conducted to evaluate EIA specificity among persons residing in Puerto Rico ( n = 534), where CM is not endemic (who were not likely to have been exposed to Coccidioides spp.), compared to blood bank donors residing in Arizona ( n = 1,218), where CM is endemic. Upon comparing serum reactivity between Puerto Rico and Arizona, the Meridian EIA showed a significant difference in IgG reactivity (0.37% versus 3.6%; P < 0.001) but not IgM reactivity (3.4% versus 2.4%; P = 0.31). No IgM-/IgG-reactive sera were detected among sera from Puerto Rico, compared to 7 (0.57%) sera from Arizona. Similar results were observed using the IMMY EIA, although significantly ( P = 0.03) fewer IgM-reactive sera from Arizona were observed, compared to the Meridian EIA. EIA-reactive sera were also evaluated by immunodiffusion before and after 3- to 4-fold concentration of the sera. These results demonstrate that elevated IgG EIA reactivity is present in sera from healthy individuals in regions of endemicity and that IgM EIA reactivity observed in sera from individuals residing outside regions of endemicity is most likely nonspecific. Other criteria, including clinical and microbiological evaluations, should be taken into account when interpreting results from surveillance studies and other reporting measures.

We report here the results of the diagnostic performances of Vitros Syphilis TPA (a chemiluminescence treponemal assay) compared with those of two treponemal enzyme immunoassays and of traditional versus reverse syphilis algorithms. Ease of use, automation, and high throughput make the Vitros Syphilis TPA assay a good choice for syphilis screening in high-volume laboratories.

Two immunoassays (Shiga Toxin Chek and Shiga Toxin Quik Chek) and real-time PCR were used to detect Shiga toxin-producing Escherichia coli . For enriched culture, the sensitivity and specificity of the three methods ranged from 80.0% to 98.2% and 98.0% to 100.0%, respectively. STEC isolates were identified in 2.6% of the 784 samples.

Serology remains critical for diagnosing hepatitis D virus (HDV) infection, which affects 15 to 20 million people worldwide, but the literature on characterizing commercial enzyme immunoassays (EIAs) dates back to 15 years ago. We evaluated 2 commercial EIAs currently available for detecting anti-HDV antibodies. The DiaSorin assay demonstrated 100% sensitivity and specificity. Using a modified cutoff value, the Cusabio assay demonstrated a sensitivity of 81.3% and specificity of 90.9%. Our data show that recently developed EIAs are reliable for anti-HDV antibody detection.

Two commonly used norovirus enzyme immunoassays have reduced diagnostic performance, with clinical sensitivities ranging from 11% to 35% for the detection of the recently emerging genogroup II genotype 17 (GII.17) Kawasaki 2014 variant that caused the majority of infections in Asia during the winter of 2014 to 2015. False-negative results can compromise infection control and patient management.

The Bio-Rad Geenius HIV 1/2 assay was evaluated as an alternative to the INNO-LIA HIV 1/2 assay for the confirmation of HIV infection in 198 serum samples reactive to 4th-generation HIV enzyme immunoassays (EIAs). The Geenius assay correctly identified 85% of the samples, compared to 75% identified by the INNO-LIA assay, reduced the number of indeterminate results, and shortened the overall turnaround time.

Following a laboratory audit, a significant number of Treponema pallidum particle agglutination assay (TPPA)-negative sera were identified when TPPA was used as a confirmatory assay of syphilis enzyme immunoassay (EIA) screening-reactive sera (SSRS). Sera giving such discrepant results were further characterized to assess their significance. A panel of 226 sera was tested by the Abbott Murex ICE Syphilis EIA and then by the Newmarket Syphilis EIA II. TPPA testing was performed on 223 sera. Further testing by the Venereal Disease Research Laboratory (VDRL) test, the Mercia Syphilis IgM EIA, the fluorescent treponemal antibody (FTA-ABS) assay, and INNO-LIA immunoblotting was undertaken in discrepant cases. One hundred eighty-seven of 223 (83.8%) SSRS were TPPA reactive, while 26 (11.6%) sera which were reactive in both the ICE and Newmarket EIAs were nonreactive by TPPA. The majority (68%) of the TPPA-discrepant sera were from HIV-positive patients and did not represent early acute cases, based on previous or follow-up samples, which were available for 22/26 samples. FTA-ABS testing was performed on 24 of these sera; 14 (58.3%) were FTA-ABS positive, and 10 (41.7%) were FTA-ABS negative. Twenty-one of these 26 sera were tested by INNO-LIA, and an additional 4 FTA-ABS-negative samples were positive. In this study, significant numbers (18/26) of SSRS- and TPPA-negative sera were shown by further FTA-ABS and LIA (line immunoblot assay) testing to be positive. The reason why certain sera are negative by TPPA but reactive by treponemal EIA and other syphilis confirmatory assays is not clear, and these initial findings should be further explored.

Various automated chemiluminescence immunoassay (CLIA) analyzers for the detection of antibodies to hepatitis C virus (HCV) are now commercially available in clinical laboratories and are replacing conventional enzyme immunoassays. We investigated the performance of four anti-HCV CLIAs (the Architect Anti-HCV assay on the Architect i2000 system, the Vitros Anti-HCV assay on the Vitros ECiQ Immunodiagnostic System, the Access HCV Ab PLUS assay on the UniCel DxI 800 analyzer, and the newly developed Elecsys Anti-HCV assay on the Cobas e 411 analyzer). The total percent coefficient of variation values of imprecision were 3.5 to 5.7% with positive control materials and 7.2 to 10.2% with negative control materials. The agreement between the results of the Elecsys, Architect, Vitros, and Access CLIAs ranged from 94.5 to 98.1%. The clinical sensitivity of all CLIAs was 100%. Each CLIA showed excellent reproducibility and clinical sensitivity. The Elecsys, Architect, Vitros, and Access CLIAs showed clinical specificities of 98.2, 98.8, 96.5, and 98.2%.

Data are accumulating on the performance of enzyme immunoassays (EIAs) for the detection of herpes simplex virus type 2 (HSV-2) infection in North America and Europe, but little is known about their performance in other populations. Nine test kits were evaluated with 330 serum samples from sub-Saharan Africa. The tests were first compared to the monoclonal antibody (MAb) EIA (Central Public Health Laboratory, London, United Kingdom). Samples that gave discordant results in the MAb EIA and in the three tests that performed best compared to the MAb EIA were tested by Western blotting (University of Washington, Seattle). A random sample of concordant samples was also tested, and the sensitivities and specificities of the different tests were calculated, taking into account this sampling strategy. The sensitivities of the tests ranged from 86 to 100%; the specificities ranged from 47 to 99%. The tests that performed best were the Gull Premier EIA (sensitivity, 86.3%; specificity, 97.6%) and the Kalon Biological (sensitivity, 92.3%; specificity, 97.7%) and Biokit (sensitivity, 86.7%; specificity, 92.6%) tests. It cannot be assumed that enzyme immunoassays for the detection of HSV-2 infection that perform well in industrialized countries will perform equally well in other populations.

Sera from 1,238 Vietnamese women in Hanoi were tested for herpes simplex virus type 2 (HSV-2). HSV-2 prevalence was 2.0%. The Kalon and Biokit assays showed significantly higher concordance to Western blotting data than did the Focus assay ( P < 0.01). Screening by Focus and then retesting with Kalon/Biokit of positive samples can reduce falsely positive results significantly ( P < 0.01).

This study describes the performance of two rapid enzyme immunoassays, Premier E. coli O157 and Premier EHEC (Meridian Diagnostics Inc., Cincinnati, Ohio) for the detection in stools of Escherichia coli O157 and verotoxins (Shiga-like toxins), respectively. Both tests were performed on stools from 876 children presenting to eight emergency departments with diarrhea. Standard culture, including E. coli O157:H7 isolation, was performed, and paired sera were taken for anti-O157-lipopolysaccharide antibody determination. Stools from patients enrolled in the study, and those yielding discordant results, were sent to a reference laboratory for repeat testing and further investigation, including cytotoxicity and non-O157 verotoxin-producing E. coli culture. Results were classified as field results (obtained in the eight site laboratories) and resolved results (obtained after repeat testing in the central laboratory). The “gold standard” for sensitivity of both tests and for specificity of Premier E. coli O157 was isolation of E. coli O157:H7 or a fourfold anti-O157 antibody rise. Specimens positive by the Premier EHEC test and negative for E. coli O157 culture were examined for non-O157 verotoxin-producing E. coli . The field sensitivity of Premier E. coli O157 was 86%, that of Premier EHEC was 89%, and the specificity of Premier E. coli O157 was 98%. Ten of 13 discordant Premier E. coli O157 results were reassigned as true results after repeat testing. Ten non-O157 verotoxin-producing E. coli isolates were recovered from Premier EHEC-positive, E. coli O157 culture-negative stools. Only one specimen gave an unequivocally false-positive Premier EHEC result. Both tests are highly sensitive and are specific if correctly performed. The Premier EHEC test will be particularly valuable as a practical routine test for the detection of non-O157 verotoxin-producing E. coli.

We evaluated four tests for the detection of rubella virus-specific immunoglobulin M antibodies. Primarily, consecutive serum samples were tested by two different assays. Selected panels of sera from patients with proven or likely recent rubella and false-positive and true-negative results in the two primary assays were further tested with two recently developed, fully automated techniques. The four tests were comparable in overall accuracy, but their dynamic ranges may differ considerably. Ways to optimize the predictive values are discussed. We conclude that automated assays may be used without causing significant changes in diagnostic accuracy or distortions in notifications of the incidence of rubella compared with the use of established tools.

The two assays evaluated in this study (the Ridascreen rotavirus and the Pathfinder rotavirus) exhibited comparable sensitivities (100%) but highly divergent positive predictive values (93.74 and 57.7%, respectively) when compared on 393 specimens. This difference should be considered when using these tests on collectives with an unknown or low prevalence.

Detection of Giardia and Cryptosporidium in clinical stool specimens using the ColorPAC and ProSpecT enzyme immunoassays revealed 98.7% agreement for Giardia detection and 98.1% agreement for Cryptosporidium detection. Sensitivities were uniformly 100%. The specificities of the ColorPAC immunoassay for Giardia and Cryptosporidium detection were 100 and 99.5%, respectively, and those for the ProSpecT assay were 98.4 and 98.6%, respectively. The false-positive reactions with the ProSpecT assay occurred with specimens that were grossly bloody.

The Meridian ImmunoCard (IC), GenBio ImmunoWELL-IgM, and Remel EIA commercial antibody tests are qualitative enzyme immunoassays that detect antibodies to Mycoplasma pneumoniae in serum. These tests were compared to an M. pneumoniae complement fixation (CF) assay, which uses a commercially available antigen component. The Meridian IC and the ImmunoWELL-IgM detect immunoglobulin M (IgM) only; the Remel EIA and the CF test detect both IgM and IgG antibodies. Detection of specific IgM antibody, which appears early in infection, can be, but is not always, indicative of a recent or current infection. Paired serum samples from 64 adult patients with probable M. pneumoniae infection were examined with each of the four tests. Thirty (47%) of the 64 acute-phase sera were IgM positive by Meridian IC, 26 (41%) were positive by Remel EIA, 24 (38%) were positive by CF, and 15 (23%) were positive by ImmunoWELL-IgM. When both the acute- and convalescent-phase serum samples from each patient were examined, 61 (95%) of the 64 patients were positive by CF, 60 patients (94%) were positive by Remel EIA, 52 patients (81%) were IgM positive by the Meridian IC, and 29 patients (45%) were IgM positive by the ImmunoWELL-IgM assay. The Meridian IC was more sensitive than the other tests for early detection of IgM antibodies. However, after examining paired serum samples, we concluded that the detection of IgM alone may not be useful for all cases of mycoplasma infection, especially in an adult population.

The use of rapid tests for human immunodeficiency virus (HIV) has become standard in HIV testing algorithms employed in resource-limited settings. We report an extensive HIV rapid test validation study conducted among Ugandan blood bank donors at low risk for HIV infection. The operational characteristics of four readily available commercial HIV rapid test kits were first determined with 940 donor samples and were used to select a serial testing algorithm. Uni-Gold Recombigen HIV was used as the screening test, followed by HIV-1/2 STAT-PAK for reactive samples. OraQuick HIV-1 testing was performed if the first two test results were discordant. This algorithm was then tested with 5,252 blood donor samples, and the results were compared to those of enzyme immunoassays (EIAs) and Western blotting. The unadjusted algorithm sensitivity and specificity were 98.6 and 99.9%, respectively. The adjusted sensitivity and specificity were 100 and 99.96%, respectively. This HIV testing algorithm is a suitable alternative to EIAs and Western blotting for Ugandan blood donors.

To determine suitability for national serosurveys, we compared two commercial enzyme-linked immunosorbent assays (ELISAs) for mumps antibody, Enzygnost Anti-Parotitis-Virus/IgG (which uses a whole-virus antigen) and Microimmune Mumps IgG Screen ELISA (which uses a recombinant nucleoprotein antigen), by testing 1,915 opportunistically collected sera submitted to diagnostic laboratories across Australia in 1997 to 1998. The proportion of positive results increased with age in both ELISAs but was significantly higher with the Microimmune than with the Enzygnost ELISA overall (88% versus 63%; P < 0.01) and in all age groups. However, the proportion of equivocal results was significantly higher with the Enzygnost than with the Microimmune ELISA (9% versus 4%; P < 0.01). Of the 572 sera with discrepant or equivocal results, 508 had sufficient sample remaining to perform the neutralization test (NT). A proportion with concordant results in both ELISAs were also tested by the NT. For sera with discrepant results, there was significantly better agreement between the NT and Microimmune than between the NT and Enzygnost (310/444 [70%] versus 135/348 [39%]; P < 0.01). Of 64 sera with equivocal Microimmune results, 45 (70%) were positive in the NT compared with 140 of 160 (88%) equivocal Enzygnost results ( P < 0.01). Compared with the NT, the Microimmune ELISA is more sensitive (96% versus 80%) but apparently less specific (36% versus 85%) than the Enzygnost ELISA. However, this is likely to be due to the generally lower sensitivity of the NT, since the Microimmune results reflect expected seroprevalence, based on vaccine uptake in the age groups studied. We conclude that the Microimmune ELISA is a more appropriate assay than the Enzygnost ELISA for estimation of mumps seroprevalence.

Highly sensitive and specific, quantitative assays are needed to detect varicella-zoster virus (VZV) immunoglobulin G in human sera, particularly for determining immune status and response following vaccination. A time-resolved fluorescence immunoassay (TRFIA) has been developed, and its performance was compared to that of two commercial enzyme immunoassays (EIAs) and Merck glycoprotein EIA (gpEIA). The TRFIA had equivalent sensitivity (97.8%) and high specificity (93.5%) in relation to gpEIA. A commercial (Behring) EIA compared favorably with TRFIA in terms of sensitivity (98.4%) but had lower specificity (80.7%). Another commercial EIA (Diamedix) had high specificity (97.1%) but low sensitivity (76.4%) compared to TRFIA if equivocal test results were treated as negative for VZV antibody. A novel feature of the TRFIA was that the cutoff was generated using population mixture modeling and was expressed in mIU/ml, as the assay was calibrated using the British standard VZV antibody.

Cobas Core Rubella IgG EIA recomb (Roche), a new, commercially available rubella enzyme immunoassay using recombinant rubella-like particles, was compared with a hemagglutination inhibition assay (Rubenosticon; Organon Teknika) and two whole-virus enzyme immunoassays (IMx [Abbott] and Platelia [Sanofi Diagnostics Pasteur]). Compared with those of these reference assays, the relative sensitivities of Cobas Core Rubella IgG recomb were 100, 94, and 95.9%, with specificities amounting to 80.8, 98, and 98.2%, respectively.

Newly available assays offer alternatives to conventional microscopic examination for Cryptosporidium spp. We compared two enzyme immunoassays, ProSpect Cryptosporidium microtiter assay (Alexon, Inc., Mountain View, Calif.) and Color Vue Cryptosporidium assay (Serady, Indianapolis, Ind.), and a direct immunofluorescent assay, Merifluor Cryptosporidium kit (Meridian Diagnostics, Cincinnati, Ohio), with acid-fast Kinyoun-staining for the detection of Cryptosporidium spp. Examinations were performed on 129 stool specimens received from patients during a recent waterborne outbreak. A specimen was considered positive when organisms could be identified visually by acid-fast and immunofluorescent stains or if organisms could be visualized by either acid-fast or immunofluorescent stain and detected by both enzyme immunoassays. The final number of positive specimens was 55. No single procedure detected all 55 positive specimens. Of these, ProSpect and Color Vue detected 52 (sensitivity, 94.5%), and the Kinyoun stain and Merifluor detected 53 (sensitivity, 96.4%). The final number of negative specimens was 74. One false-positive result was seen with both the Kinyoun stain and the ProSpect assay. The Color Vue and ProSpect assays required the most hands-on technologist time. The ProSpect assay and Merifluor kit were easiest to perform. The acid-fast stain was difficult to interpret. The Merifluor kit was easiest to read and was adaptable to both batch and single testing. Overall, the Kinyoun stain and the Merifluor test were preferable to both of the enzyme immunoassays because of the high reagent cost and hands-on time required for the enzyme immunoassays. The difficult interpretation of the Kinyoun stain smears made the Merifluor a more desirable test despite its higher cost. We conclude that all methods tested were equally sensitive and specific for the detection of Cryptosporidium spp. Ease of use, adaptability to batch testing, and cost are important criteria in determining the method of choice.

We evaluated the performance of three commercial measles immunoglobulin M enzyme immunoassays from Meddens, Denka Seiken, and Behring. The sensitivities were determined to be 96.7% for the Meddens and Denka Seiken assays and 87.9% for the Behring assay. The specificities of the assays were determined to be 94.6% for Meddens, 98.2% for Denka Seiken, and 98.7% for Behring.

We read with interest the paper “Evaluation of the C.Diff Quik Chek Complete Assay, a New Glutamate Dehydrogenase and A/B Toxin Combination Lateral Flow Assay for Use in Rapid, Simple Diagnosis of Clostridium difficile Disease” by Sharp and colleagues (10). Interest in the laboratory diagnosis of Clostridium difficile has been encouraged by research that has shown the commonly used toxin enzyme immunoassays (EIAs) to be inadequate when used alone (1, 9). We agree with Sharp et al. (10) that glutamate dehydrogenase (GDH) is a very sensitive (but poorly specific) assay and can accurately rule out the presence of C. difficile in stool samples. Sharp and colleagues (10) used a commercially available combined GDH and toxin A/B assay (C.Diff Quik Chek Complete) in the first step of their testing algorithm, as others have done (2, 8, 11). However, we note that there were no true-positive samples that were GDH negative but EIA toxin positive in the 284 specimens they tested. We made the same observation in our own study of 500 specimens (3), and others have reported similar results (6, 7, 11). Thus, it is very rare to have a GDH-negative, EIA toxin-positive result for a true-positive sample. Therefore, we believe the toxin component of the C.Diff Quik Chek Complete assay is redundant. We prefer to use the GDH-only assay (C.Diff Chek-60), which is less expensive and allows for automated processing using the Dynex DS2 platform. This saves hands-on time and avoids possible misinterpretation of ambiguous results, since it produces a numerical result. We note that Sharp and colleagues had one false-positive result with their testing algorithm, giving a sensitivity and specificity of 100% and 99.6%, respectively (10). Using those authors' figures, we calculated performance characteristics using our preferred algorithm of screening with the C.Diff Chek-60 GDH assay, followed by confirmation with GeneXpert PCR, and found a sensitivity and specificity of 100% and 100%, respectively. Furthermore, the GeneXpert system can simultaneously indicate the presence of the potentially “hypervirulent” ribotype 027 strain (4), giving important epidemiological information not provided by the C.Diff Quik Chek Complete test. Using the Sharp and colleagues' figures for material cost per test (10), we also calculated the cost of each algorithm for testing 284 specimens. Those authors suggested the algorithm would cost $4,200.64, and ours cost slightly less at $3,923.30 (Table ​(Table1).1). Turnaround times for these algorithms would not differ significantly (depending upon degree of batching), but significant reductions may be possible by using GeneXpert as a point-of-care test. Further study is warranted in this area. TABLE 1. Material cost comparison for two suggested diagnostic algorithms It is clear that the GDH test is sufficiently sensitive to work well as an initial screen, but we do not believe any benefit is derived by coupling it with a toxin EIA. Furthermore, a dedicated, automated GDH test is easier to interpret. In England, we are now seeing a decrease in the prevalence of Clostridium difficile infection (which is a mandatory reportable disease) (5), and this will inevitably have an impact upon the positive and negative predictive values of any diagnostic tests. It is critical, therefore, that we have robust, accurate methods of identifying patients with this organism.

The efficacy of pneumococcal vaccines in protecting against pneumococcal pneumonia can feasibly be measured only with a diagnostic technique that has a high specificity (0.98 to 1.00) and a sensitivity greatly exceeding that of blood cultures (>0.2 to 0.3). In this context immune-complex enzyme immunoassays (EIAs) offer a novel, convenient diagnostic method, and we have investigated three such assays with appropriate study populations in Kenya. Sera from 129 Kenyan adults with pneumococcal pneumonia and 97 ill controls from the same clinics, but without pneumococcal disease syndromes, were assayed with immune-complex EIAs for pneumolysin, C-polysaccharide, and mixed capsular polysaccharides (Pneumovax II). At an optical density (OD) threshold yielding a specificity of 0.95, the sensitivities (95% confidence intervals) of the assays were 0.22 (0.15 to 0.30), 0.26 (0.19 to 0.34), and 0.22 (0.15 to 0.29), respectively. For pneumolysin immune complexes, human immunodeficiency virus (HIV)-positive patients had a higher mean OD than HIV-negative patients (639 versus 321; P < 0.0001), but stratification by HIV infection status did not alter the performance of this test. Combining the results of all three EIAs did not enhance the diagnostic performances of the individual assays. In Kenyan adults the sensitivities of the immune-complex EIAs could exceed that of blood cultures only at levels of specificity that were insufficient for the performance of vaccine efficacy studies.

Rapid detection of group A rotavirus was performed by using ImmunoCardStat! Rotavirus (ICS-RV) (which uses immunogold-based, horizontal-flow membrane technology), two commercial enzyme immunoassays (Premier Rotaclone and TestPack Rotavirus), and electron microscopy. A total of 249 stool specimens collected from children with gastroenteritis between February and April 1997 were tested. After resolution of 19 of the 22 discordant results by reverse transcription-PCR for group A rotavirus, ICS-RV detected 125 positives while Rotaclone and TestPack detected 127 and 129 positives, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value were 94.0, 100, 100, and 93.4% for ICS-RV; 95.5, 100, 100, and 95.0% for Rotaclone; and 97.0, 96.5, 97.0, and 96.5% for TestPack. ICS-RV was sensitive and specific and was relatively simple to perform and interpret.

Galactomannan (GM) detection by the Platelia Aspergillus enzyme immunoassay is widely used for diagnosing invasive aspergillosis. A major problem with the detection of circulating GM is the occurrence of false-positive results due to the cross-reactivity of the test with antigens other than

Test strips for the detection of antibodies to human immunodeficiency virus type 1 were investigated using specimens from risk groups in Thailand (141 reactive; 445 nonreactive) in a local Thai laboratory. The diagnostic sensitivity and specificity were both 100%. Using a set of seroconversion panels, the sensitivity of the test strips was within the range of sensitivities obtained with enzyme immunoassays. The test was developed for performance at decentralized settings under nonlaboratory conditions.During December 1992-January 1993, the Thai Ministry of Public Health collected blood samples from populations at high risk of HIV infection (prostitutes and men attending sexually transmitted disease clinic). All these samples were evaluated for HIV infection by enzyme immunoassay and, if positive, confirmed by the Western blot. There were 141 HIV-1 reactive samples. In a blind study, laboratory personnel of the Venereal Diseases and Control Center in Chonburi, Thailand, later analyzed 586 serum samples with test strips (Sero-Strip HIV; Saliva Diagnostics Systems; Singapore). The strips have a synthetic peptide representing a highly conserved region of viral transmembrane glycoprotein, gp41. This peptide captures HIV-1 antibodies. Three investigators read the test strips independently. There were no discrepant readings among them. The test strips were also evaluated under nonlaboratory conditions of decentralized settings: exposure for 21 weeks at 45 degrees Celsius, exposure for 20 weeks at 22 degrees Celsius, and 98% relative humidity. The results of the test strips correlated completely with those of the conventional immunoassays. Thus, there were neither false positive nor false negative reactions. In conclusion, these test strips can be used reliably in non-air-conditioned tropical settings.

We noted with interest a recent letter (G. Giocoli, Letter, J. Clin. Microbiol. 38:3520–3521, 2000) exhorting microbiologists to report likelihood ratios (LRs) along with diagnostic test results. We wish to expand on the strengths and weaknesses of LRs for infectious disease diagnosis. As Giocoli points out, sensitivity and specificity reflect the probability of a test result when disease status is known, whereas for individual patients we wish to know the probability of disease given the test result. With a physician's estimate of the pretest probability, LRs allow estimation of disease probability given a test result—seemingly the ideal diagnostic parameter. LRs may be calculated from 2×2 tables or directly from the sensitivity (Se) and the specificity (Sp) (2): LR for positive result = (Se)/(1 − Sp); LR for negative result = (1 − Se)/(Sp). LRs allow estimation of a test's contribution to decision-making. A clinician estimates the pretest probability of disease, either from prevalence data alone or in combination with clinical risk factors. A positive diagnostic test increases the likelihood of the diagnosis, whereas a negative test makes it less likely. LRs quantitate how much the test result has altered the disease probability. The posttest probability should be high enough to confirm the diagnosis or low enough to rule out disease. If the posttest probability falls between these two thresholds, further testing is required. The clear advantage of LRs is to permit the calculation of posttest probabilities with the help of a convenient nomogram (Likelihood Ratios; Center for Evidence-Based Medicine [http://cebm.jr2.ox.ac.uk/docs/likerats.html]), unlike sensitivity and specificity, which require conversion to prevalence-corrected predictive values. Furthermore, LRs can be applied to more than two outcomes of a test (2). For example, enzyme immunoassays often define a positive cutoff. LRs may be applied to different levels of cutoffs, such that a high optical density is associated with a higher LR (and hence is more likely to confirm a diagnosis) than with a low, but still positive, result. However, clinicians rarely use LRs. They have difficulty estimating pretest probabilities, do not carry nomograms, and disagree on thresholds for ruling out, or ruling in, disease. In general, clinicians claim to prefer sensitivity and specificity, albeit with their own clinical experience, as the “gold standard” for assessing these parameters (1). LRs are summary measures which enable microbiologists to better express diagnostic test performance and facilitate clinician estimation of posttest probabilities of disease. We believe that LRs are simpler and more intuitively used than sensitivity and specificity and will lead to improved test utilization and decision-making through an understanding of the clinical impact of a test result. Until there is more general acceptance and use of LRs as summary parameters, the reporting of LRs for positive and negative test results should complement, rather than supplant, sensitivity and specificity.

The efficacies of nine enzyme-linked immunosorbent assays (EIA) for antibody to human immunodeficiency virus type 1 (HIV-1) and one EIA for antibody to HIV-2 in detecting antibody to HIV-2 were studied. The competitive EIAs for antibody to HIV-1 were less sensitive than the indirect EIAs. The overall prevalence of positive results was between 28 and 51% with the competitive EIAs and between 70 and 93% with the indirect EIAs. Most of the EIAs were less sensitive in detecting antibody to HIV-2 in sera from people with acquired immunodeficiency syndrome-like diseases than in sera from symptomless individuals. The results indicate that there is a high degree of cross-reactivity between HIV-1 and HIV-2 by EIA, indicating that serotype specificity must be determined by Western blot (immunoblot) with both sets of viral antigens. The results are relevant for discussing public health strategies, especially the screening of blood donors; competitive EIAs for antibody to HIV-1 are not sensitive enough to be used in areas where HIV-2 is prevalent (West Africa).The efficacy of 10 enzyme immunoassays (EIAs) for antibody to human immunodeficiency virus type 1 (HIV-1) in detecting antibody to HIV-2 was evaluated in sera from 43 West African residents. 23 sera were from asymptomatic individuals belonging to high-risk groups (prostitutes, prisoners, patients with sexually transmitted diseases), while the remaining 20 serum samples were from persons with symptoms of acquired immunodeficiency syndrome (AIDS). The overall prevalence of positive results was 28-51% with the competitive EIAs and 70-93% with the indirect EIAs. Most of the EIAs were less sensitive in detecting antibody to HIV-2 in sera from people with AIDS-like disease compared with asymptomatic individuals. This study shows that commercial HIV-1 antibody test kits are not equally efficient in detecting HIV-2-antibody-positive sera. Cross-reacting HIV-2 antibodies seem not to have enough affinity to compete with the anti-HIV-1 conjugate. Given the high degree of cross-reactivity between HIV-1 and 2 by EIA, serotype specificity must be determined by Western blot with both sets of viral antigens. In areas such as the Ivory Coast where HIV-2 is prevalent, these findings must be taken into consideration in the screening of blood donors.

The diagnosis of congenital cytomegalovirus (CMV) infection is often accomplished by the detection of circulating antibody directed against CMV. We devised a method for measuring CMV-specific immunoglobulin M (IgM) based on the isolation of IgM antibody by reaction with a solid phase coated with antihuman IgM. The determination of IgM antibody specific for CMV was accomplished by the subsequent addition of CMV or control antigen and enzyme-labeled CMV antibody (solid phase-IgM method). We compared the sensitivity and specificity of this method with those of a conventional form of solid-phase enzyme immunoassay in which CMV antigen is bound to the solid phase (solid phase-antigen method). Both assay systems were capable of detecting CMV-specific IgM antibody in the sera of 10 babies with documented CMV infection and in those of the mothers of 4 of these babies. The solid phase-IgM method yielded negative results in all 66 sera available from babies who did not have congenital CMV infection. On the other hand, the solid phase-antigen system yielded false-positive results in 12 (18%) of these sera. In addition, the solid phase-antigen system yielded false-positive results in 8 of 12 sera obtained from patients with demonstrable rheumatoid factor. However, the solid phase-IgM system yielded negative results for the rheumatoid sera, provided that appropriate control reactions were performed. The solid phase-IgM system is thus a specific and sensitive method for the determination of CMV IgM antibody.

Une infection par le virus HDV (hepatitis delta virus) est determinee par la detection d'un antigene de ce virus et d'un anticorps dirige contre ce virus. Deux methodes de detection avaient ete utilisees par Dubois et Goudeau: Dettassay (Noctech) et la technique immunoenzymatique Organon, le premier s'etant revele moins sensible que le second. Or, Shattock et Morris ont des resultats inverses. L'explication de ce phenomene reside probablement dans la matrice des reactifs utilises pour ces deux tests. En effet, le test Organon mis en pratique par Dubois et Goudeau a ete modifie par rapport au test d'origine. Les divergences de resultats peuvent donc s'expliquer de cette maniere