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1.
Pak J Med Sci ; 40(2ICON Suppl): S28-S34, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38328653

ABSTRACT

Background & Objective: Nearly 80 million of the Pakistani population received two doses of the BBIBP-CorV vaccine, against SARS-CoV-2, and 2.6 million people received heterologous booster doses up to February 2022. Our objective was to measure the long-term change of antibody titers in persons vaccinated with Pfizer-BioNTech COVID-19 following two doses of BBIBP-CorV. Methods: Serum specimens from forty-three participants were collected 4-8 weeks following two doses of BBIBP-CorV at the Indus Hospital & Health Network, Karachi. A second set of serum specimens were collected 2-12 months after Pfizer-BioNTech COVID-19 booster dose administration. Chemiluminescent Microparticle Immunoassay (CMIA, Abbott Alinity Quant), and the pseudotyped lentivirus antibody neutralization assay were performed on all specimens. The latter assay was reported as log half-maximal inhibitory concentrations (IC50), calculated using a nonlinear regression algorithm (log [inhibitor] versus normalized response variable slope) in Graph Pad Prism 9. Paired sample t-test was used to ascertain the statistical significance of the difference in means of antibody titers obtained before and after the booster vaccine doses. Results: Mean log10 values obtained with CMIA before and after the booster dose were 2.90 AU/mL and 3.87 AU/mL respectively, while the corresponding log10 IC50 values obtained through pseudotyped lentivirus antibody neutralization assay were 2.45 and 2.80. These differences were statistically significant with CMIA (p = <0.00001), but not with pseudotyped lentivirus antibody neutralization assay (p = 0.06318.). Conclusion: A heterologous booster dose with Pfizer-BioNTech COVID-19 vaccine following two doses of BBIBP results in increased total antibody titers, though neutralizing antibody titers may start to wane a few months after the booster dose.

2.
Pract Lab Med ; 36: e00326, 2023 Aug.
Article in English | MEDLINE | ID: mdl-37649536

ABSTRACT

Objective: To verify the analytical performance of cobas® HBV PCR and cobas® HCV PCR assays with Abbott m2000 RealTime System as the reference method. Design: De-identified residual, archived patient specimens, and College of American Pathologists (CAP) proficiency testing samples were used. Analytical parameters verified were accuracy, precision, limit of detection (LOD), linear range, and cross-contamination. Experiments were designed in accordance with Clinical Laboratories Standards Institute (CLSI) guidelines and CAP standards. Analysis of accuracy was done through regression plots and Bland Altman analyses. Precision was analyzed through coefficient of variation and ANOVA; LOD through probit analysis; and linear range through polynomial fit analysis. Results: The regression plots for accuracy showed a slope nearing 1, with a y-intercept close to zero, while Bland Altman analyses also showed no systematic evidence of bias, though concordance of results was not perfect near the lower limit of quantification. Coefficients of variation were all below 15%, while ANOVA returned p-values above 0.99, indicating no statistically significant imprecision. The LOD verified were an order of magnitude higher than the manufacturer reported ones for both assays, while the linear range verified was more limited. Within the verified range, polynomial fit analysis showed line to be the best fit for the data. Conclusions: cobas® HBV PCR and cobas® HCV PCR assays showed acceptable accuracy, acceptable precision, as well as no evidence of cross-contamination. The LOD verified were higher, and linear ranges more limited than those reported by the manufacturer. Verifications of these may be limited by availability of appropriate testing specimens.

3.
J Clin Virol Plus ; 3(1): 100137, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36644775

ABSTRACT

With widespread global COVID-19 vaccine coverage, a scalable, cost-effective, and standardized tool to ascertain post-vaccine immunity is a dire need. Neither clinical evaluations of vaccine efficacy, nor live virus antibody neutralization assays fulfill these criteria. Commercially available anti-S binding immunological assays have the potential to fill this gap, but need to be systematically evaluated for their utility to serve as surrogates for the aforementioned, widely accepted tools of determining vaccine efficacy. In this study, we evaluated an anti-S binding immunological assay (Roche Elecsys Anti-SARS-CoV-2 S) by utilizing two hundred and fifty-five archived serum specimens, either pre-pandemic, or those exposed to natural infections or vaccines with their neutralizing titers pre-determined through a live virus, pseudotyped antibody neutralization assay. Roche Elecsys Anti-SARS-CoV-2 S demonstrated good sensitivity (98%) and specificity (99%), just as has been reported in some other previously conducted studies using this assay. Only a mild correlation, however, with the live virus pseudotyped lentivirus antibody neutralization assay (Spearman's r = 0.26) was observed. We conclude that, as such, Elecsys Anti-SARS-CoV-2 S has a high sensitivity and specificity for detecting anti-SARS-CoV-2 S proteins, though the assay does not always correlate well with live virus assays for quantitative outcomes.

4.
Vaccines (Basel) ; 10(5)2022 Apr 28.
Article in English | MEDLINE | ID: mdl-35632448

ABSTRACT

Fifty five percent of the Pakistani population is still unvaccinated with the two-dose protocol of COVID-19 vaccines. This study was undertaken to determine the seroconversion rate and antibody titers following the two-dose BBIBP-CorV protocol, and to compare these variables in unvaccinated, COVID-19 recovered individuals (total n = 180) at Indus Hospital and Health Network, Karachi. Pseudotyped lentivirus antibody neutralization assays and SARS-CoV-2 IgG Quant II (Abbott) immunoassays were performed 4-8 weeks following the second dose of the BBIBP-CorV or PCR positivity/onset of symptoms of COVID-19. Seroconversion rate, using neutralization assays, in vaccinated individuals was lower (78%) than that in unvaccinated, COVID-19-recovered individuals with moderate to severe infection (97%). Prior PCR positivity increased serocoversion rate to 98% in vaccinated individuals. Immunoassays did not, however, reveal significant inter-group differences in seroconversion rates (≥95% in all groups). Log10 mean antibody neutralizing titers following the two-dose BBIBP-CorV protocol (IC50 = 2.21) were found to be significantly less than those succeeding moderate to severe COVID-19 (IC50 = 2.94). Prior SARS-CoV-2 positivity significantly increased post-vaccination antibody titers (IC50 = 2.82). Similar inter-group titer differences were obtained using the immunoassay. BBIBP-CorV post-vaccination titers may, thus, be lower than those following natural, moderate to severe infection, while prior SARS-CoV-2 exposure increases these titers to more closely approximate the latter.

5.
Glob Health Sci Pract ; 10(1)2022 02 28.
Article in English | MEDLINE | ID: mdl-35294380

ABSTRACT

As the coronavirus disease (COVID-19) pandemic spread, meeting the testing needs to control the spread of infection became a major challenge worldwide. In Pakistan, the lack of the requisite infrastructure and training compounded the acute shortage of testing kits and other consumables. Against this backdrop and to urgently improve province-wide access to high-quality COVID-19 polymerase chain reaction (PCR) testing with rapid turnaround times, the Government of the Sindh (GoS) province of Pakistan entered into a public-private partnership with Indus Hospital & Health Network (IHHN). Under this partnership, the GoS undertook sample collection and Indus Hospital in Karachi, Sindh, centralized testing. We describe the implementation strategies adopted by the partnership, as well as the challenges, opportunities, and lessons learned. Notably, up to 40% and 22% of total COVID-19 PCRs done in Sindh in the first 2 months of the pandemic, respectively, were performed at Indus Hospital in Karachi, though this percentage declined gradually as other centers caught up with their testing capacities. The rapid scaling up was achieved through a combination of mechanisms and factors including building on preexisting partnerships between the GoS and IHHN, pooling resources and harnessing distinct and complementary roles, relocating existing resources, introducing automation and information technology system changes, establishing risk mitigation strategies, and introducing quality measures within testing processes. The primary outcome of the partnership was rapid province-wide access to quality COVID-19 PCR testing with short turnaround times and at no cost to the patient. Furthermore, implementation of the partnership goals established new mechanisms as well as strengthened existing ones to enable rapid response to the future global health security challenges in Sindh, Pakistan.


Subject(s)
COVID-19 , Public-Private Sector Partnerships , COVID-19/diagnosis , COVID-19/epidemiology , COVID-19 Testing , Global Health , Humans , Pakistan/epidemiology
6.
Pak J Med Sci ; 38(2): 393-398, 2022 Jan.
Article in English | MEDLINE | ID: mdl-35310800

ABSTRACT

Objective: To determine the stability of respiratory samples for SARS-CoV-2 PCR at standard laboratory ultra-freezer temperatures (-80°C). Methods: Five hundred and sixty-five archived, SARS-CoV-2 PCR positive patient specimens received at the Pathology Department of the Indus Hospital & Health Network between January 2021 and June 2021 were retested in June 2021. Samples had been stored at -70°C or below throughout this duration. Sample integrity following storage was assessed as the percentage of samples with reproducible results, and as consistency of cycle threshold (Ct) values between the original testing and the repeat testing. Results: Of the 565 samples evaluated in this study, 86% gave reproducible results upon retesting. However, there was no correlation between the duration of storage and result reproducibility, though the majority (69% for PCR Target-I and 78% for PCR Target-II respectively) of non-reproducible results had Ct values above 30. Similarly, there was a consistent increase of Ct values upon storage at ultra-freezer temperatures, though the effect again was more contingent upon freezing the sample in the ultra-freezer rather than the duration of storage. Conclusion: SARS-CoV-2 positive respiratory specimens for PCR can be stored for up to six months at -70°C or below without loss of sample integrity, though there is some loss of PCR-detected viral targets as evidenced by an immediate increased in the PCR-generated Ct values. In addition, samples with initial Ct values above 30 are more likely to give non-reproducible results.

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