Optical immunoassay test for rapid detection of influenza A and B viruses: an evaluation

The optical immunoassay test (FLU OIA, BioStar, USA) for rapid detection of influenza A and B viral antigens was compared with viral isolation in cell culture. A total of 103 respiratory specimens were tested on 75 pediatric patients with acute respiratory illnesses. Influenza viruses were recovered in 40 specimens (type A: 5, Type B: 35). FLU OIA demonstrated 80.0% sensitivity and 68.8% specificity for nasopharyngeal aspirates and 36.7% sensitivity and 83.9% specificity for throat swabs. We also tested FLU OIA, retrospectively, using 78 supernatant samples from pediatric patients with influenza A virus infection frozen after cell culture. FLU OIA demonstrated 91.4% sensitivity and 92.3% specificity for nasopharyngeal aspirates and 50.0% sensitivity and 91.7% specificity for throat swabs diluted in viral transport media. Nasopharyngeal aspirates showed higher sensitivity than throat swabs for detection of influenza virus by FLU OIA. We believe this rapid test kit is useful for the detection of influenza A and B viruses.     

Evaluation of an immunochromatography test using enzyme immunoassay for rapid detection of influenza A and B viruses

We evaluated the performance of an improved version of Espline Influenza A & B-N (Fujirebio Inc., Japan), an immunochromatography test using enzyme immunoassay for rapid diagnosis of influenza A and B. The test produced positive results for four strains of influenza viruses and thirty-one influenza viral antigens and negative results for all of thirty strains of other respiratory viruses that were tested. The detection limit of this test was 5.8 x 10(2) to 5.8 x 10(3) pfu/assay, which is more sensitive than the old version of Espline. Furthermore, 715 respiratory specimens collected from the patients (children, 79.4%; adults, 18.5%; unknown, 2.1%) with influenza-like illnesses during the 2002/2003 influenza season in Japan were tested as part of a clinical evaluation of this test. The relative performance of this test compared to cell culture and nested RT-PCR results were examined. In the cell cultures, influenza viruses were detected in 488 of the 715 specimens (overall, 68.3%; AH3, 41.7%; B, 26.4%; AH3 and B, 0.1%). For influenza A, the sensitivity of this test was 95.4% (125/131) for nasal aspirates, 96.8% (92/95) for nasal swabs, and 85.1% (63/74) for throat swabs. For influenza B, the sensitivity of this test was 91.2% (52/57) for nasal aspirates, 88.1% (59/67) for nasal swabs, and 71.6% (48/67) for throat swabs. The new test exhibited a remarkably higher sensitivity to influenza A in throat swabs than the old version of Espline. Only two false positive results were obtained out of a total of 223 virus negative specimens; the specificity of the test was 100% (88/88) for nasal aspirates, 97.6% (81/83) for nasal swabs, and 100% (52/52) for throat swabs. We conclude that the new Espline Influenza A&B-N rapid diagnostic test is easy to use and has a high sensitivity and specificity, especially for influenza A.     

Evaluation of the rapid detection test for influenza A and B viruses using neuraminidase activity

The ZstatFlu test (ZymeTx, USA) is a rapid detection kit for influenza A and B viruses. This test is based upon the reaction between viral neuraminidase from influenza viruses and a chromogenic substrate. The clinical performance of the ZstatFlu test was determined by comparison with viral isolation in cell culture. A total of 176 respiratory specimens from 172 pediatric patients with influenza like illnesses during the 1998/99 season were tested. Influenza viruses were recovered from 97 specimens (type A: 6, type B: 91) in cell culture. ZstatFlu demonstrated 67.4% sensitivity (29/43) and 62.7% specificity (37/59) for throat swabs. Of the 22 ZstatFlu-positive, culture-negative throat swabs tested by RT-PCR, 18 were positive by RT-PCR. ZstatFlu showed 48.1% sensitivity (26/54) and 90.0% specificity (18/20) for nasopharyngeal aspirates. Of the two ZstatFlu-positive, culture-negative nasopharyngeal aspirates tested by HI titer of paired sera, one showed a 4-fold increase of HI titer. Nasopharyngeal aspirates therefore showed lower sensitivity than throat swabs at this test, different from EIA test kits such as Directigen FluA or FLU OIA. Overall, only 5 specimens were false positive by the ZstatFlu test. Therefore, this test demonstrated high specificity and positive predictive value. In conclusion, the ZstatFlu test is useful for the rapid detection of influenza A and B viruses to identify patients who need antiviral treatment.     

Evaluation of immunochromatography method for rapid detection of influenza A and B viruses

We have evaluated a new rapid detection kit for influenza A and B viruses, known as the QuickVue Influenza test (Quidel Coporation, USA); which is based on immunochromatography using virus isolates and clinical specimens. Twelve strains of influenza A and B were tested for evaluate the reactivity and detection limits of this test. The QuickVue Influenza test showed a positive result for all twelve strains of influenza virus and a negative result for fourteen different kinds of other respiratory viruses. The detection limits for six strains were 5 to 30 pfu/ml for a cell culture, 1.0 x 10(3) to 6.0 x 10(4) pfu/ml for 1st PCR, 1 to 50 pfu/ml for nested PCR, 3.0 x 10(5) to 6.0 x 10(5) pfu/ml for the QuickVue Influenza test, 1.5 x 10(5) to 1.0 x 10(6) pfu/ml for the Directigen Flu A, and 7.5 x 10(5) to 5.0 x 10(6) pfu/ml for the FLU OIA. Furthermore, the QuickVue Influenza test were clinically evaluated using 92 throat swab specimens collected from patients with influenza-like illnesses. By cell culture, influenza viruses were detected in 49 of the 92 specimens (AH1N1: 20, AH3N2: 7, B: 22); the titers of the influenza viruses were between 2.5 pfu/ml and 7.0 x 10(5) pfu/ml. Compared to cell culture, the QuickVue Influenza test showed a sensitivity of 75.5%, a specificity of 93.0%, a positive predictive value of 92.5%, a negative predictive value of 76.9%, and an efficiency value of 83.7%. On the other hand, influenza viruses were detected in 54 of the 92 specimens (AH1N1: 19, AH 3N2: 10, B: 25) by RT-PCR. Compared to RT-PCR, the QuickVue Influenza test showed a sensitivity of 72.2%, a specificity of 97.4%, a positive predictive value of 97.5%, a negative predictive value of 71.2%, and an efficiency value of 82.6%. Overall, only one throat swab specimen produced a false positive result using the QuickVue Influenza test; thus, this test appears to have a high specificity. We conclude that the QuickVue Influenza test is a simple one-step test with a sensitivity and specificity equivalent to those of other conventional diagnostic kits. The test is useful and suitable for the diagnosis of influenza and for identifying influenza patients requiring antiviral therapy.     

Sensitivity and specificity of rapid influenza testing of children in a community setting

Please cite this paper as: Stebbins et al. (2011) Sensitivity and specificity of rapid influenza testing of children in a community setting. Influenza and Other Respiratory Viruses 5(2), 104–109. Introduction Rapid influenza testing (RFT) allows for a rapid point‐of‐care diagnosis of influenza. The Quidel QuickVue^® Influenza A+B test (QuickVue) has a reported manufacturer’s sensitivity and specificity of 73% and 96%, respectively, with nasal swabs. However, investigators have shown sensitivities ranging from 22% to 77% in community settings. Methods The QuickVue rapid influenza test was evaluated in a population of elementary (K‐5) school children, using testing in the home, as part of the Pittsburgh Influenza Prevention Project during the 2007–2008 influenza season. The QuickVue test was performed with nasal swab in full accordance with package instructions and compared with the results of nasal swab semi‐quantitative RT‐PCR. Results Sensitivity of the QuickVue was found to be 27% in this sample. There was no statistically valid correlation between the semi‐quantitative PCR result and the QuickVue result. Conclusions This study is consistent with the low sensitivity of the QuickVue test also reported by others. Viral load, technique, and the use of nasal swabs were examined as contributing factors but were not found to be explanations for this result. Community testing includes patients who are on the lower spectrum of illness which would not be the case in hospital or clinic samples. This suggests that RFT is less sensitive for patients at the lower spectrum of illness, with less severe disease.     

Comparison of three rapid diagnostic kits using immunochromatography for detection of influenza A virsuses

In 2004, 3 new rapid influenza diagnostic kits using immunochromatography that allow type differentiation became commercially available. They are the ESPLINE Influenza A & B-N (Fujirebio Corp., Japan: ESPLINE-N hereafter), QuickVue Rapid SP influ (Quidel Corp., USA: QuickVue), and POCTEM INFLUENZA A/B (INTERNATIONAL REAGENTS Corp., Japan: POCTEM). The authors performed a prospective study that compared the usefulness among the 3 kits in 151 children with suspected influenza, who were examined within 3 days after onset, between January and March, 2004. Nasopharyngeal aspirates were collected, and viruses were isolated. The residual samples were diluted and centrifuged, and the supernatant was used for the rapid diagnosis tests. Influenza virus AH3 was isolated in 95 children and influenza B virus in 3. In the 95 children with influenza virus AH3, the sensitivity and specificity of ESPLINE-N were 100% and 100%, respectively, those of QuickVue were 99% and 91%, and those of POCTEM were 91% and 100%. The sensitivity of POCTEM was significantly lower than that of the other 2 kits (p < 0.01), and the specificity of QuickVue was significantly lower than that of the other 2 kits (p < 0.05). Examination was performed within 1 day after onset in 55 of the 95 children, including 30 who underwent examination within 6 hours after the development of fever. The body temperature was less than 38.0 degrees C in 14 of the 95 children. In all children including these children, virus detection was possible by ESPLINE-N. ESPLINE-N allowed very accurate diagnosis of influenza A using samples prepared by diluting and centrifuging nasopharyngeal aspirates.     

Clinical evaluation of rapid diagnostic kit detecting separately influenza A and B viruses

The Directigen Flu A + B kit, a rapid diagnostic device for influenza virus A and B was evaluated. The nasopharyngeal aspirates were obtained from 239 patients who visited our hospital, between January and March, 2000, presenting flu-like symptoms. Influenza virus AH1: 77 and AH3: 51 were isolated from 128 specimens and none from 111 specimens. Directigen Flu A + B showed 115 specimens positive and 106 specimens negative. The sensitivity and specificity of this kit were 89.8% (115/128) and 95.5% (106/111) compared with viral isolation. Agreement on positive and negative interpretations between Direction Flu A and this kit was 97.9% (234/239). In the evaluation of this kit for influenza B virus, 60 frozen nasopharyngeal aspirates collected from February to April, 1999 were used. The sensitivity and specificity of this kit were 88.9% (16/18) and 88.1% (37/42) compared with viral isolation. Agreement on positive and negative interpretations between FLU OIA and this kit was 91.7% (55/60). The Directigen A + B demonstrated sensitivity and specificity equivalent to the conventional kits in nasopharingeal aspirates. This kit can also differentiate influenza A and B viruses, a feature which is useful for treatment using anti-viral agents such as amantadine and neuraminidase inhibitor. To date, the kit is the most effective tool for the rapid diagnosis of influenza.     

Evaluation of an immunochromatography test kit for rapid diagnosis of influenza

We assessed the sensitivity and specificity of the Capilia Flu AB rapid diagnosis kit for influenza that utilizes the immunochromatography method. Tested were 114 influenza like illness patients in the 2001/2002 influenza season. We used Capilia Flu AB and Infu A . B Quick, a rapid diagnosis kit based on enzyme immunoassay. As laboratory confirmation tests, influenza virus isolation and polymerase chain reaction (PCR) were done. Those patients with positive results from virus isolation or PCR were regarded as influenza patients. The sensitivities of nasal swab, pharyngeal swab, and nasal wash specimens were 82.8%, 80.0%, and 75.0%, respectively. The specificities of nasal swab, pharyngeal swab, and nasal wash specimens were 95.3%, 93.9%, and 100%, respectively. A total of 20 patients displayed different results in comparison of their nasal and pharyngeal swabs: 15 patients were positive with the nasal swab but negative with the pharyngeal swab and 5 patients were negative with the nasal swab but positive with the pharyngeal swab. Nasal swab would seem to be preferable in terms of sensitivity. The sensitivity and specificity of Capilia Flu AB were a little higher than those of Influ A . B Quick, but with no significant difference. The one-step operation of Capilia Flu AB is easier than the four steps required by Influ A . B Quick, but the time required to make a diagnosis is the same. No significant age related difference in the effectiveness of the kits was found. The Capilia Flu AB rapid diagnosis kit is useful in clinical practice because it has good sensitivity (about 80%) and specificity (about 90%), and it is easy to use.     

Are current rapid detection tests for Group A Streptococci sensitive enough? Evaluation of 2 commercial kits

A new, 1-step, enzyme-linked immunoassay kit for detection of Group A Streptococci (GAS) in throat samples (QuickVue In-Line One-Step Strep A Test; Quidel Corporation, San Diego, CA) was evaluated for use in a study comprising 536 patients in 8 primary healthcare centres. Compared to conventional culture at the clinical microbiology laboratory, the sensitivity achieved was 73.9% and the specificity 86.8%; these figures were not affected to any major extent by broth enrichment of samples before culturing or following PCR testing of the cysteine proteinase gene for independent diagnosis of GAS. It was also found that most samples containing low numbers of GAS were missed by the rapid test. We therefore evaluated the kit in use in our area (TestPack Plus Strep A Test; Abbott Laboratories, Chicago, IL) in a separate study of 615 patients. Somewhat increased sensitivity (82.8%) and specificity (96.1%) were obtained. As current antigen tests depend on subjective judgement of test outcome, improvements in test design or provision of more detailed instructions may be desirable in order to achieve optimal results.     

Evaluation of direct and rapid identification of group A streptococci from throat swabs by Culturette 10-Minute Group A Strep ID test kit

The Culturette Brand 10-Minute Group A Strep ID test kit (Marion Scientific, Division of Marion Laboratories, Inc., Kansas City, Mo.) was evaluated for its sensitivity and specificity in identifying the group A streptococci directly from 96 throat swabs, against the conventional culture method and serological grouping test. Our results indicated that the rapid test kit and conventional method are 93.8% accurate; percent sensitivity and specificity of the rapid test was 80.6% and 100%, respectively. None of the false-positive observed in rapid test kits occurred with the heterogeneous microorganisms. More than 8 X 10(4) colony forming unit per swab was required for the positive latex agglutination of the test kit. Since the Culturette method is simple to perform and correctly identifies group A streptococcal antigen, and also required no special instruments, it appears to be applicable in hospital laboratories and outpatients clinics.     

Clinical performance of three rapid diagnostic tests for influenza virus in nasopharyngeal specimens to detect novel swine-origin influenza viruses

Background  

Influenza is an important public health problem. The aim of this study was to evaluate and compare the sensitivity and specificity of three rapid diagnostic tests (SEKISUI, QuickVue Influenza A + B, and SD BIOLINE) for novel swine-origin influenza viruses (S-OIV) and seasonal influenza.     

Use of a rapid detection assay for influenza virus, on nasal aspirate specimens

We investigated the usefulness of a rapid antigen detection kit using optical immunoassay for influenza virus (FLU OIA, BioStar, USA). Nasal aspirates were taken from 92 influenza suspected outpatients between March to April of 1999. Compared with virus isolation and PCR, the sensitivity of FLU OIA was 88.5% and 81.6%, and the specificity was 65.2% and 72.2%. All isolated viruses were influenza type B virus. It was difficult to differentiate the weak-positive and negative cases, leading to the rather low specificity, although the assay procedure was easy and quick. FLU OIA may be a useful rapid diagnosis kit for influenza in pediatric outpatient clinics and wards, because it can detect both influenza type A and type B viruses.     

Comparison of four rapid diagnostic kits using immunochromatography to detect influenza B viruses

We compared the usefulness of 4 rapid influenza diagnostic 1-device kits using immunochromatography, which facilitate type differentiation, i.e. ESPLINE Influenza A&B-N (Fujirebio Corp., Japan: ESPLINE), POCTEM INFLUENZA A/B (Sysmex Corp., Japan: POCTEM), Quick Vue Rapid SP influ (Quidel Corp., U.S.A.: Quick Vue), and Capilia Flu A + B (TAUNS Corp., Japan: Capilia), in 278 children in whom influenza infection was suspected in 2004 and 2005. Nasopharyngeal aspirates were diluted for virus isolation and residual samples were centrifuged. Using the supernatant, we conducted rapid diagnosis testing. Influenza virus AH3 was isolated from 40 children, and influenza B virus from 163. Of the 40 children, the sensitivity and specificity of ESPLINE, POCTEM, Quick Vue, and Capilia were 100%/100%, 95%/100%, 98%/96%, and 98%/96%. In the 163 children, the sensitivity and specificity were 89%/100%, 87%/100%, 88%/97%, and 86%/98%. ESPLINE showed the highest sensitivity and specificity to influenza viruses AH3 and B. All kits were less sensitive to influenza B virus than to influenza A virus, however. The specificity of Quick Vue and Capilia was low; so these kits must be improved.     

Influenza surveillance in Pune, India, 2003

Influenza surveillance was conducted in Pune, India in 2003. A total of 573 throat swabs/ nasal swabs (TS/NS) and 190 nasopharyngeal aspirates (NPA) were collected from 763 in- and out-patients who were mostly children aged 0-16 years. TS/NS (507/573) and NPA (42/190) specimens were processed in MDCK cell cultures and identified with the hemagglutination inhibition test (HI). A total of 37 influenza viruses was isolated: twenty-three type A (H3N2) and 14 type B of the Yamagata lineage were isolated from 29 children and 8 adults. Three type A (H3N2) isolates were characterized as being similar to A/Panama/2007/99 like, A/Korea/770/2000 like, and B/Sichuan/379/99 like strains.     

一种A型流感病毒NP抗原快速检测试剂的建立

目的建立一种适合现场检测需要的A型流感快速诊断试剂。方法以自制的抗A型流感(FluA)病毒核蛋白(NP)单抗为原料,建立快速检测A型流感病毒的双抗体夹心酶免疫渗滤试验,并对其灵敏度和特异性进行了初步评价。结果该试剂对不同地区流行的各种亚型的A型流感病毒株均有较高的反应性,而对非FluA病毒株无交叉反应。比较该试剂与BD公司的两种流感快速诊断试剂,发现该试剂对随机选取的3株FluA病毒的检测分析灵敏度高出Directigen EZ Flu A试剂5~125倍,对2株FluA病毒的分析灵敏度高出Directigen Flu A试剂约20倍。另外,用该试剂对57份含漱液标本和170份动物拭子标本进行检测,结果显示:本试剂的灵敏度(>85%)和特异性(>95%)均优于当前主流的商品化A型流感快速诊断试剂。结论利用抗FluANP单抗为原料建立了A型流感快速诊断试剂,该试剂的应用无需任何专用仪器,操作简便快速,可满足现场检测需要。     

Evaluation of Wondfo influenza A&B fast test based on immunochromatography assay for rapid diagnosis of influenza A H1N1

Influenza viruses cause significant morbidity and mortality in both children and adults during local outbreaks or epidemics. Therefore, a rapid test for influenza A&B would be useful. This study was conducted to evaluate the clinical performance of the Wondfo influenza A&B test for rapid diagnosis of influenza A H1N1 Infection. The rapid testing assay could distinguish infection of influenza A and B virus. The reference viral strains were cultured in MDCK cells while TCID50 if the viruses were determined. The analytical sensitivity of the Wondfo kit was 100 TCID50/ml. The Wondfo kit did not show cross reactivity with other common viruses. 1928 suspected cases of influenza A (H1N1) virus infection were analyzed in the Wondfo influenza A&B test and other commercially available products. Inconsistent results were further confirmed by virus isolation in cell culture. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 100%, 98.23%, 92.45%, and 100% for flu A, and 96.39%, 99.95%, 98.77%, and 99.84% for flu B respectively. 766 suspected cases of influenza A (H1N1) virus infection were analyzed in the Wondfo influenza A&B test and RT-PCR. The sensitivity, specificity, PPV and NPV were 56.5%, 99.75%, 99.52% and 71.04% for flu A, 25.45%, 99.86%, 93.33% and 94.54% for flu B respectively. These results indicate that the Wondfo influenza A&B test has high positive and negative detection rates. One hundred fifty-six specimens of influenza A (H1N1) confirmed by RT-PCR were analyzed by the Wondfo influenza A&B test and 66.67% were positive while only 18.59% were positive by the reference kit. These results indicate that our rapid diagnostic assay may be useful for analyzing influenza A H1N1 infections in patient specimen.     

Study of a respiratory syncytial virus diagnostic test kit based on immunochromatography

We carried out clinical and basic studies of the Directigen Lateral Flow RSV (Becton, Dickinson and Company, USA), a rapid test kit that detects respiratory syncytial virus (hereinafter referred to as "RSV") antigens based on immunochromatography. For the clinical study, 103 nasopharyngeal aspirates from patients with acute respiratory infections were used to evaluate the kit. Compared to the cell culture method, the Directigen Lateral Flow RSV showed a sensitivity of 100% (16/16) and a specificity of 94.3% (82/87), and an agreement rate of 95.1% (98/103). When compared to conventional testing kits, we found that the total agreement rate with the Directigen RS (Nippon Becton Dickinson and Company) was 88.3% (91/103) and with RSV TestPack (Dainabot Co., Ltd.) was 91.3% (94/103). The detection limit of the Directigen Lateral Flow RSV was 2 x 10(3) PFU/ml for both RSV subgroups A and B. In the crossreactivity test, only RSV was found positive. No other microorganisms were crossreactive. We also studied storage stability of nasopharyngeal aspirates and found that stability was not affected by storage at room and refrigerator temperatures for 14 days. Taken all together, the Directigen Lateral Flow RSV is useful for the diagnosis of RSV infection in a clinical setting because its performance is equivalent to conventional testing kits and is easy to use.