Chi Hyun Cho

Evaluation of five rapid diagnostic kits for influenza A/B virus

Influenza viruses cause seasonal epidemics associated with high morbidity and mortality. However, even during periods of epidemic prevalence, clinical diagnoses are problematic. Rapid diagnostic tests for the detection of pandemic influenza A/B virus are valuable for their ease of use. Many rapid influenza diagnostic kits were introduced recently in the Republic of Korea (ROK), including Directizen EZ Flu A and B (Becton Dickinson, Sparks, USA), Binax Now Influenza A/B antigen kit (Binax, Portland, USA), Genedia influenza Ag (Green Cross, Yongin, ROK), Humasis Influenza A/B antigen test (Humasis, Anyang, ROK), and SD Bioline rapid influenza kit (Standard Diagnostics, Yongin, ROK). The objective of this study was to evaluate the performance of these five rapid diagnostic kits. The results were compared with those of viral culture and reverse transcription (RT)-PCR. A total of 253 nasopharyngeal swabs were analyzed from 253 patients (influenza A, n=67; B, n=86; negative samples, n=100). The specimens were tested immediately by conventional influenza virus culture and RT-PCR, stored at -80°C, and tested using five rapid test kits. The performance of the five rapid tests kits varied with sensitivities between 71.0 and 82.1% and between 37.2 and 47.7% for detecting influenza A and B, respectively. For influenza A, the sensitivities of the Directizen EZ Flu A and B, Binax Now Influenza A/B antigen kit, Genedia influenza Ag, Humasis Influenza A/B antigen test, and SD Bioline rapid influenza kits were 82.1%, 71.0%, 76.1%, 79.1%, and 82.1%, respectively; those for influenza B were 40.7%, 37.2%, 40.7%, 41.8%, and 47.7%, respectively. The specificity of all rapid tests was 100%. Commercial influenza antigen detection assays are useful tools for the rapid diagnosis of influenza. However, confirmatory testing is always recommended.

Evaluation of Sofia fluorescent immunoassay analyzer for influenza A/B virus

BACKGROUND The influenza virus causes seasonal epidemics which are associated with high morbidity and mortality. Rapid diagnostics tests (RDT) are frequently used to make a quick influenza diagnosis to confirm the clinical suspicion, despite their low sensitivity. OBJECTIVES Assess the performance of the Sofia Influenza A+B Fluorescence Immunoassay (Quidel, San Diego, CA). STUDY DESIGN Nasopharyngeal swabs, taken from 241 patients (influenza A (n=73)/B (n=72), negative samples (n=96)) were analyzed using the Sofia Influenza A+B Fluorescence Immunoassay, BinaxNOW Influenza A/B antigen kit (Alere Inc., USA), Directigen EZ Flu A and B (Becton Dickinson, USA), real-time RT-PCR and an influenza virus culture. RESULTS There was a significant difference between the performance of rapid antigen tests and the Sofia FIA, when compared to the RT-PCR, in the detection of influenza strain A and B. Indeed, the Sofia FIA displayed sensitivities of 82.2% and 77.8% for strains A and B respectively, whereas sensitivities of BinaxNOW Influenza A/B antigen kit, and Directigen Flu A and B were 54.8%, and 68.5% for influenza A, and 62.5%, and 52.8% for influenza B respectively. The average RT-PCR threshold cycle (C(t)) (±SD) for the Sofia Influenza A+B Fluorescence Immunoassay-positive specimens was higher than those of the BinaxNOW Influenza A/B antigen and the Directigen EZ Flu A and B kit positive specimens. CONCLUSION Compared to other RDTs, the Sofia Influenza A+B Fluorescence Immunoassay is a sensitive, and rapid method for the detection and discrimination between influenza A and B.

Evaluation of a novel real-time RT-PCR using TOCE technology compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses

Abstract Background Various kinds of commercial molecular systems have been developed for fast and more accurate detection of respiratory viruses. Anyplex™ II RV16 [RV16] was designed for simultaneous detection of 16 respiratory viruses using multiplex PCR coupled with TOCE™ technology. Objectives To compare the performance of RV16 with those of culture and Seeplex® RV15 ACE [RV15] by determining their sensitivity and specificity. Study design Seven hundred and thirty respiratory samples were tested by modified shell vial culture method, RV16, and RV15. For molecular tests, automated nucleic acid extraction and liquid handling system using MICROLAB Nimbus IVD (Hamilton, USA) was adopted to maximize the workflow and accuracy. Performance of each assay was determined against a composite reference standard. Results Two hundred and one samples (28%) out of 730 samples were positive by culture, while additional 281 (39%) were positive by RV16 or RV15. Sensitivities of RV16, RV15, and culture for virus tested were as follows: 100/93/63% for influenza A, 90/80/69% for influenza B, 98/94/63% for RSV, 98/52/23% for adenovirus, and 100/75/46% for PIV. For viruses not covered by culture, sensitivities of RV16 and RV15 were as follows: 99/81% for rhinovirus, 92/100% for coronavirus OC43, 100/56% for coronavirus 229E/NL63, 92/88% for metapneumovirus, 100/62% for bocavirus, and 91/91% for enterovirus. Overall, the specificities of culture, RV16, and RV15 (Seegene) were 100/99.9/99.9%. Conclusions RV16 assay was superior to culture method and RV15 and will be a promising tool for patient management and public health epidemiology.

Evaluation of the AdvanSure™ real-time RT-PCR compared with culture and Seeplex RV15 for simultaneous detection of respiratory viruses

Abstract Recently, AdvanSure™ kit based on multiplex real-time PCR was developed for simultaneous detection of 14 respiratory viruses (RVs). We compared the performance of AdvanSure with those of Seeplex® RV 15 ACE and culture by determining their sensitivities and specificities against a composite reference standard. Four hundred thirty-seven respiratory samples were tested by modified shell vial culture method, RV 15 ACE, and AdvanSure. One hundred fourteen samples (26.2%) out of 437 samples were positive by culture, while additional 91 (20.8%) were positive by AdvanSure or RV15. One hundred twelve of 114 culture-positive samples were positive by AdvanSure except 2 samples (1 adenovirus, 1 respiratory syncytial virus [RSV]). Overall, the sensitivities of culture, RV15, and AdvanSure were 74.5%, 89.8%, and 95.1%, respectively. Sensitivities of culture, RV15, and AdvanSure for each virus tested were as follows: 91/100/96% for influenza A, 60/0/100% for influenza B, 63/95/97% for RSV, 69/81/89% for adenovirus, and 87/93/93% for parainfluenza virus. For viruses not covered by culture, sensitivities of RV15 and AdvanSure were as follows: 77/88% for rhinovirus, 100/100% for coronavirus OC43, 40/100% for coronavirus 229E/NL63, 13/100% for metapneumovirus, and 44/100% for bocavirus. The overall specificities of culture, RV15, and AdvanSure were 100/98.9/99.5%, respectively. Of 45 coinfected specimens, AdvanSure detected 41 specimens (91.1%) as coinfected, while RV15 detected 27 specimens (60.0%) as coinfected. AdvanSure assay demonstrated exquisite performance for the detection of RVs and will be a valuable tool for the management of RV infection.

Clinical performance evaluation of the BD Veritor System Flu A+B assay.

Early identification of influenza is important for optimal patient management and infection control. Rapid influenza antigen tests have been used routinely in clinical settings to confirm clinical suspicion, despite their low sensitivity. To improve sensitivity, various influenza point-of-care test reader systems have been developed. This study evaluated the clinical performance of a digital readout rapid influenza diagnostic test (RIDT), the BD Veritor™ System Flu A+B assay (BD). Nasopharyngeal swabs taken from 250 patients (influenza A positive, n=75; influenza B positive, n=75; and influenza negative, n=100) were analyzed using the BinaxNOW® Influenza A/B antigen kit (BN), SD Influenza Ag A/B kit (SD), BD, real-time reverse transcriptase polymerase chain reaction (RT-PCR), and an influenza virus culture. Compared to RT-PCR, the sensitivities of BN, SD, and BD were 56.0, 53.3, and 72.0%, respectively, for influenza A and 57.3, 65.3, and 69.3%, respectively, for influenza B. No false-positive results were noted with the three rapid antigen tests. For influenza A, the average RT-PCR threshold cycle (Ct) for specimens that tested positive using BD was higher than that for specimens that tested positive using BN and SD. BD is a sensitive and easy method for the early detection of influenza A and B.

Genetic variability in Plasmodium vivax aldolase gene in Korean isolates and the sensitivity of the Binax Now malaria test

Introduction of rapid malaria diagnostic tests (RDT) initiated numerous field evaluations in various epidemiologic settings. But the efficiency of some RTD kits based on aldolase raised reservations for direct implementation of RDT into clinical settings. We performed Binax Now malaria test in 84 Korean Plasmodium vivax isolates and compared it with the traditional Giemsa stain microscopy test as the reference standard. The sensitivity of Binax Now was 62.0% for P. vivax cases (52/84, 95% CI 51.2–71.6%) with 100.0% specificity (50/50, 95% confidence interval 92.9–100%). After the aldolase gene sequence analysis of 84 isolates, two synonymous mutations in aldolase gene were identified in both Binax Now positive and negative samples. No significant association between the mutations and Binax Now malaria tests was found. Thus, the genetic variability would not explain the poor performance of P. vivax RDTs by detecting aldolase in ROK isolates.

Drilling and deburring in a single process

Abstract A drill bit integrated with a retractable deburring cutter can be used for drilling and deburring exit burrs in a single process. In preliminary works, the authors learned that the retractable cutter may cause hole widening during its passage through the holes. A new design for the deburring cutter is proposed to avoid hole widening. With the new design, mild exit burrs (types I and II) are successfully removed. For large exit burrs (type III), however, the deburring is likely to be incomplete.

Comparison between Saliva and Nasopharyngeal Swab Specimens for Detection of Respiratory Viruses by Multiplex Reverse Transcription-PCR

ABSTRACT Nasopharyngeal swabs (NPSs) are being widely used as specimens for multiplex real-time reverse transcription (RT)-PCR for respiratory virus detection. However, it remains unclear whether NPS specimens are optimal for all viruses targeted by multiplex RT-PCR. In addition, the procedure to obtain NPS specimens causes coughing in most patients, which possibly increases the risk of nosocomial spread of viruses. In this study, paired NPS and saliva specimens were collected from 236 adult male patients with suspected acute respiratory illnesses. Specimens were tested for 16 respiratory viruses by multiplex real-time RT-PCR. Among the specimens collected from the 236 patients, at least 1 respiratory virus was detected in 183 NPS specimens (77.5%) and 180 saliva specimens (76.3%). The rates of detection of respiratory viruses were comparable for NPS and saliva specimens (P = 0.766). Nine virus species and 349 viruses were isolated, 256 from NPS specimens and 273 from saliva specimens (P = 0.1574). Adenovirus was detected more frequently in saliva samples (P < 0.0001), whereas influenza virus type A and human rhinovirus were detected more frequently in NPS specimens (P = 0.0001 and P = 0.0289, respectively). The possibility of false-positive adenovirus detection from saliva samples was excluded by direct sequencing. In conclusion, neither of the sampling methods was consistently more sensitive than the other. We suggest that these cost-effective methods for detecting respiratory viruses in mixed NPS-saliva specimens might be valuable for future studies.

Clinical Performance Evaluation of the Sofia RSV FIA Rapid Antigen Test for Diagnosis of Respiratory Syncytial Virus Infection

ABSTRACT A recently introduced Sofia respiratory syncytial virus (RSV) fluorescent immunoassay (FIA) was evaluated against the BinaxNOW RSV card and the SD Bioline RSV test using 348 respiratory samples. The Sofia, BinaxNOW, and SD Bioline kits showed sensitivities of 66%, 65%, and 64%, respectively, for detecting RSV-A, and 71%, 63%, and 65% for detecting RSV-B, respectively.

Evaluation of Plasmodium vivax ELISA for the blood screen

Plasmodium vivax malaria is the indigenous strain in the Republic of Korea (ROK). Plasmodium vivax can be transmitted through the transfusions of various blood components, which became a severe problem with the safety of blood transfusions and blood‐related products in ROK. We evaluated a P. vivax‐specific enzyme‐linked immunosorbent assay (Genedia Malaria Ab ELISA 2.0, Green Cross, ROK) with blood samples from four groups: 251 samples from P. vivax‐infected patients, 39 samples from post‐treatment patients upon follow‐up, 200 samples from healthy volunteers and 421 samples from domestic travellers to and from high endemic areas of ROK. The positive cases from the ELISA test were confirmed by both Giemsa microscopic and polymerase chain reaction methods. The clinical sensitivity and specificity of detecting P. vivax with ELISA test were 94.4% and 99.0%, respectively. Thirteen of 421 domestic travellers (3.0%) to endemic areas tested positive. The results indicate the effectiveness of detecting antibodies against P. vivax in blood with Genedia Malaria Ab ELISA 2.0 test in a large blood screen setting.