Evaluation of a novel anterior nasal swab for the detection of SARS-CoV-2.

BACKGROUND: During the course of the COVID-19 pandemic, nasopharyngeal swabs, combined throat and nose swabs and saliva samples have been evaluated for SARS-CoV-2 detection using nucleic acid amplification tests (NAT). Literature on anterior nasal swabs is limited. We investigated a novel anterior nasal swab that has been designed to standardised self-collection, maximise sample uptake and improve user comfort. We used combined throat and nose swabs and neat saliva samples as the comparators. RESULTS: The overall positive percentage agreement between the Rhinoswab™ and the combined throat and nose swab was 95.2 % at day 2 post participant recruitment and 93.3 % on day 4 post participant recruitment. This was favourable to the positive percentage agreement with saliva at the same time points. CONCLUSION: In our study the Rhinoswab™ performed equally well in comparison to a combined throat and nose swab for the laboratory detection of SARS-CoV-2 using nucleic acid amplification techniques.

Trends in Clostridioides difficile diagnosis before and after a change in testing algorithm.

Clostridioides difficile (Clostridium difficile) (CD) infection remains a challenging diagnosis in hospitalized patients given the myriad of testing procedures and array of alternative causes for diarrhea. We identified 100 consecutive inpatients with positive CD testing in a single tertiary center before and after changing from nucleic acid amplification testing (NAAT) alone to a two-step algorithm involving Glutamate Dehydrogenase enzyme immunoassays (GDHEIA) followed by an enzyme immunoassay for CD toxins (EIA). Detailed clinical information was obtained retrospectively to assess for risk factors, clinical features, and treatment outcomes to correlate test results with clinical cases. We demonstrate that using a 2-step testing algorithm identifies patients with a consistent clinical illness for CD disease significantly more often than nucleic acid amplification testing alone without an increase in cases of severe CD disease. Our data suggest that NAAT alone results in an increase in unnecessary treatment of CD colonization.

Unusual clinical presentations of new-onset herpetic eye disease after ocular surgery.

PURPOSE: To report five cases of new-onset herpetic eye disease with unusual presentation after ocular surgery. METHODS: Herpetic eye disease was suspected in five cases, three after cataract surgery and two after lamellar corneal transplantation surgery. Of these, four cases presented within 2-6 weeks of surgery. The clinical presentation was in the form of an epithelial defect, suspected epithelial down growth, graft oedema with unexplained anterior chamber inflammation and graft-host interface infection. A swab for viral detection with real-time polymerase chain reaction was performed in all the described cases. RESULTS: Herpes simplex disease was detected in all cases. All cases responded to the antiherpetic medications. CONCLUSIONS: Our study shows that new-onset herpetic eye disease may occur after cataract surgery and lamellar corneal transplantation, and a high index of suspicion may be necessary for the diagnosis in such cases.

High viral fitness during acute HIV-1 infection.

Several clinical studies have shown that, relative to disease progression, HIV-1 isolates that are less fit are also less pathogenic. The aim of the present study was to investigate the relationship between viral fitness and control of viral load (VL) in acute and early HIV-1 infection. Samples were obtained from subjects participating in two clinical studies. In the PULSE study, antiretroviral therapy (ART) was initiated before, or no later than six months following seroconversion. Subjects then underwent multiple structured treatment interruptions (STIs). The PHAEDRA study enrolled and monitored a cohort of individuals with documented evidence of primary infection. The subset chosen were individuals identified no later than 12 months following seroconversion to HIV-1, who were not receiving ART. The relative fitness of primary isolates obtained from study participants was investigated ex vivo. Viral DNA production was quantified using a novel real time PCR assay. Following intermittent ART, the fitness of isolates obtained from 5 of 6 PULSE subjects decreased over time. In contrast, in the absence of ART the fitness of paired isolates obtained from 7 of 9 PHAEDRA subjects increased over time. However, viral fitness did not correlate with plasma VL. Most unexpected was the high relative fitness of isolates obtained at Baseline from PULSE subjects, before initiating ART. It is widely thought that the fitness of strains present during the acute phase is low relative to strains present during chronic HIV-1 infection, due to the bottleneck imposed upon transmission. The results of this study provide evidence that the relative fitness of strains present during acute HIV-1 infection may be higher than previously thought. Furthermore, that viral fitness may represent an important clinical parameter to be considered when deciding whether to initiate ART during early HIV-1 infection.

Assessing proficiency of interpretation of rapid human immunodeficiency virus assays in nonlaboratory settings: ensuring quality of testing.

Rapid antibody tests for the detection of human immunodeficiency virus (HIV) offer an effective means of providing a timely result of HIV serostatus to individuals. The increased use of rapid HIV antibody tests outside the laboratory has highlighted the need for new, cost-effective quality assurance methods to be developed for use in nonlaboratory-based and resource-limited settings. Photographed rapid HIV test results were used in a modified external quality assessment scheme to assess the interpretation proficiency and, therefore, to assess the feasibility of using this method as a basis for a quality assessment program for nonlaboratory-based testing. Participants (n = 148), both experienced and inexperienced in the performance and interpretation of rapid HIV testing, interpreted the photographed results of five rapid HIV assays. These were scored according to the degree of technical discordance. Error scores were grouped according to each participant's technical experience. The accuracy of interpretation for four of the five assays was between 80 and 97%, indicating that the photographed results of samples, including those difficult to read or borderline difficult to read, can be used to assess the proficiency of test operators in interpreting results. Participants had greater difficulty in interpreting samples of weak reactivity; this was consistent across the five assays. Experience played an important role in accurate interpretation, with experienced laboratory participants exhibiting greater proficiency (P < 0.05) in interpreting the results of three of the five rapid HIV assays. It was established that photographed results of rapid HIV assays could be interpreted with accuracy and demonstrated that prior experience resulted in a more accurate interpretation performance.

The prevalence and risk behaviours associated with the transmission of blood-borne viruses among ethnic-Vietnamese injecting drug users.

OBJECTIVE: To measure the prevalence and determinants of blood-borne virus (BBV) transmission in ethnic Vietnamese injecting drug users (IDUs). METHODS: The study was conducted in Melbourne, Australia, in 2003. It was a cross-sectional design with participants recruited from street-based illicit drug markets predominately using a snowball technique. One hundred and twenty-seven participants completed a questionnaire that asked about illicit drug use and participants' blood samples were tested for HIV, HCV and HBV. RESULTS: One hundred and three (81.1%) ethnic Vietnamese IDU study participants were HCV positive and three (2.4%) were HIV positive. More than 60% had evidence of being infected with HBV (either in the past, acute infection or chronic infection). Almost 60% had injected daily over the past 12 months. Fifty-nine participants had recently travelled to Vietnam; 24 (41%) had injected drugs in Vietnam; and three (12.5%) reported sharing injecting equipment in Vietnam. CONCLUSION: The prevalence of BBVs was higher in this study's IDU population compared with IDUs in Australia generally, despite the fact that the injecting risk behaviours were similar to IDUs more generally. IMPLICATIONS: Culturally sensitive drug treatment and education programs need to be developed in Australia for both ethnic Vietnamese IDUs and their families to reduce this group's risk of contracting a BBV.

Change in hepatitis C virus genotype in injecting drug users.

Six major genotypes of the hepatitis C virus (HCV) have been described; it is assumed to be uncommon for genotypes to change in chronically infected individuals. Venous blood samples obtained from Vietnamese-Australian injecting drug users who participated in successive studies conducted in Melbourne, Australia, were genotyped using the Bayer line probe assay and genotype confirmed by sequencing whenever possible. Three changes of HCV genotype were observed, and one infection in an individual not exposed previously. The rate of change of genotype was 3 in 11.4 person-years (py), or 26.4 per 100 py (95% CI: 8.5, 81.6). Traditionally-calculated HCV incidence was 1 in 4.3 py, or 23.3 per 100 py (95% CI: 3.3, 165.1). These data imply that HCV genotype change in injecting drug users occurs at least as frequently as infections in naive individuals, and that traditionally-calculated HCV incidence rates represent a minority of actual HCV transmission among practicing injecting drug users.

Validation of assembled nucleic acid-based tests in diagnostic microbiology laboratories.

Medical microbiology and virology laboratories use nucleic acid tests (NAT) to detect genomic material of infectious organisms in clinical samples. Laboratories choose to perform assembled (or in-house) NAT if commercial assays are not available or if assembled NAT are more economical or accurate. One reason commercial assays are more expensive is because extensive validation is necessary before the kit is marketed, as manufacturers must accept liability for the performance of their assays, assuming their instructions are followed. On the other hand, it is a particular laboratory's responsibility to validate an assembled NAT prior to using it for testing and reporting results on human samples. There are few published guidelines for the validation of assembled NAT. One procedure that laboratories can use to establish a validation process for an assay is detailed in this document. Before validating a method, laboratories must optimise it and then document the protocol. All instruments must be calibrated and maintained throughout the testing process. The validation process involves a series of steps including: (i) testing of dilution series of positive samples to determine the limits of detection of the assay and their linearity over concentrations to be measured in quantitative NAT; (ii) establishing the day-to-day variation of the assay's performance; (iii) evaluating the sensitivity and specificity of the assay as far as practicable, along with the extent of cross-reactivity with other genomic material; and (iv) assuring the quality of assembled assays using quality control procedures that monitor the performance of reagent batches before introducing new lots of reagent for testing.

Anti-HCV confirmatory testing of voluntary blood donors: comparison of the sensitivity of two immunoblot assays.

BACKGROUND: In this study, the sensitivity of two commercially available anti-HCV immunoblot assays (HCV Western blot (Wellcozyme] and RIBA 3.0 SIA [RIBA-3, Chiron]) was compared in a voluntary blood donor population. STUDY DESIGN AND METHODS: Four groups of donor samples were retrospectively tested in this study. Groups 1 and 2 were donor samples that gave positive or indeterminate band patterns, respectively, when originally tested on the HCV Western blot between 1994 and 1998. These samples were tested on the RIBA 3.0. Donor samples in Groups 3 and 4 were originally tested on RIBA-3 during 1998 and 1999 and gave positive or indeterminate blot results, respectively. In this study these two groups were tested on the HCV Western blot. Samples with discrepant results on the two immunoblot assays were selected for genotyping or serotyping. RESULTS: The two immunoblots showed similar sensitivity to the core and NS5 proteins. However, of 35 samples positive on Western blot or RIBA-3, the Western blot failed to detect NS4 in 14 samples compared with only 5 for RIBA-3. As well, the Western blot failed to detect NS3 in 6 samples compared to 2 for RIBA-3. Five (27.8%) of 18 samples that were Western blot indeterminate due to core reactivity showed an additional NS3 band on RIBA-3. Of the samples with additional NS3 and/or NS4 reactivity on RIBA-3 that were genotyped or serotyped, all were HCV type 3. CONCLUSIONS: Western blot and RIBA-3 showed similar sensitivity to the HCV core and NS5 proteins. However, RIBA-3 showed greater sensitivity to both NS3 and NS4 compared to the Western blot. The reduced sensitivity of the Western blot to the NS3 and NS4 proteins was observed with HCV type 3 samples.