IgG antibody response against nucleocapsid and spike protein post-SARS-CoV-2 infection.

OBJECTIVES: Coronavirus disease-19 (COVID-19) pandemic became the greatest public health challenge globally. Study of dynamicity and durability of naturally developed antibodies against SARS-CoV-2 are of great importance from an epidemiological viewpoint. METHODS: In this observational cohort study, we have followed up the 76 individuals who tested positive for SARS-CoV-2 infection by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) for 16 weeks (post-enrolment) to record the periodic changes in titre, concentration, clinical growth and persistence of naturally developed SARS-CoV-2 antibodies. We collected serum samples from these individuals for 16 weeks with a frequency of weekly and fortnightly during each follow-up and tested them in two CLIA-based platforms (Abbott Architect i1000SR and Roche Cobas e411) for testing SARS-CoV-2 antibodies both qualitatively and quantitatively. RESULTS: We recorded the antibody magnitude of these individuals 10 times between September 2020 and February 2021. We found a waning of antibodies against nucleocapsid antigen protein but not a complete disappearance by the end of 16 weeks. Out of 76 cases, 30 cases (39.47%) became seronegative in qualitative assay, although all the sera samples (100%) remained positive when tested in quantitative assay. CONCLUSION: The lower persistence of anti-nucleocapsid SARS-CoV-2 antibody may not be the exact phenomenon as those cases were still seropositive against spike protein and help in neutralising the virus.

Pooled testing for SARS-CoV-2 infection in an automated high-throughput platform.

BACKGROUND: Active detection of SARS-CoV-2 infection through testing is elementary for the control of COVID-19 pandemic. The implementation of large-scale RT-PCR testing has led to a rise in the demand for testing kits whose availability is always a concern. OBJECTIVE: To find out the feasibility of pooled testing in a high-throughput platform. METHODOLOGY: Pooled testing was conducted in Roche cobas 6800 in 2 methods. Firstly, the simple two-stage testing algorithm was conducted for 1410 samples individually and then as pooled samples. Secondly, we evaluated the sensitivity of cobas 6800 for the detection of a single positive sample within a pool of negative samples. RESULTS: Implementing the five-sample Dorfman pooling to test 1410 samples, we identified 42 (2.9%) individual SARS-CoV-2-positive samples and 27 (9.5%) positive pool samples. The pooling strategy precisely identified all the positive samples. All individually negative samples were also accurately determined by pooling. There was 100% sensitivity of detecting positive samples in a pool of negative samples even up to 1:64 dilution. There was a threefold increase in total throughput in one-third of the cost per day. CONCLUSION: A high-throughput platform such as Cobas 6800 can effectively increase the testing capacity by twofold to threefold by adopting the pooled testing strategy for successful management of SARS-CoV-2 and helping in the containment of community transmission.

Scrub typhus seroprevalence from an eastern state of India: findings from the state-wide serosurvey.

BACKGROUND: Scrub typhus is a mite-borne infectious disease caused primarily by the obligate intracellular bacteria Orientia tsutsugamushi, which is transmitted by chigger mites. The objective of this study was to determine the prevalence of scrub typhus among adults in Odisha, an eastern state of India. METHODS: A descriptive cross-sectional study was conducted to analyse 1840 serum samples from five districts (Khordha, Cuttack, Ganjam, Malkangiri and Sundargarh) of Odisha collected during 2020-2021. Both immunoglobulin G (IgG) and IgM antibodies against scrub typhus were tested using commercial enzyme-linked immunosorbent assay kits. Point estimates of the 95% confidence interval and adjusted odds ratio were calculated. RESULTS: Of 1840 participants, the prevalence of IgG positivity was 1034 (56.19%) and that of IgM was 523 (28.42%). The majority of participants were in the 18-45 y age group (53.7%). Cuttack had the highest IgG positivity (64.54%) and Malkangiri had the lowest (29.32%). The highest and lowest positivity for IgM were found in Malkangiri (40.84%) and Cuttack (25.30%), respectively. CONCLUSIONS: With an increasing number of infections detected in the state, scrub typhus is emerging as a public health threat. Increasing awareness among the general public and healthcare professionals through health education campaigns regarding scrub typhus is essential. Early detection of the disease through the establishment of a laboratory surveillance system is required to control the transmission of scrub typhus.

Diagnostic accuracy comparison of three fully automated chemiluminescent immunoassay platforms for the detection of SARS-CoV-2 antibodies.

BACKGROUND: Serological test is an essential surveillance tool to track down the extensiveness of SARS-CoV-2 transmission and subsequently to move out from the enforced lockdown stage. OBJECTIVE: The study measures the diagnostic accuracy of three popular chemiluminescent immunoassay (CLIA) based automated platforms for the detection of anti-SARS-CoV-2 antibodies and compares their agreements. STUDY DESIGN: Serum samples of 594 COVID-19 positive patients and 100 samples from pre-COVID cases were tested by three CLIA based automated platforms: Abbott architect i2000SR, Roche cobas e411 and Yhlo iFlash 1800 and their diagnostic accuracy were compared by the area under the curves (AUC) value obtained from receiver operator characteristic (ROC) curves. Cohen's kappa statistic and McNemar's test were used to interpret the agreement between the platforms. RESULTS: All three platforms showed high specificity as claimed by the manufacturer. Sensitivity was calculated as 64.48 % (58.67-70.3) for Abbott, 80.48 % (76.62-84.34) for Roche and 76.94 % (72.65-81.23) for Yhlo. AUC was maximum for Roche (0.929). The Cohen's kappa value was determined in between 0.69-0.89 as the inter-rater agreements. CONCLUSION: The overall statistical analysis demonstrated cobas e411 as the diagnostically most accurate platform among the three.

SARS-CoV-2 IgG antibody responses in rt-PCR-positive cases: first report from India.

INTRODUCTION: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody responses remain poorly understood and the clinical utility of serological testing is still unclear. AIM: To understand the relationship between the antibody response to SARS-CoV-2 infection and the demographics and cycle threshold (C t) values of confirmed RT-PCR patients. METHODOLOGY: A total of 384 serum samples were collected from individuals between 4-6 weeks after confirmed SARS-CoV-2 infection and tested for the development of immunoglobulin class G (IgG) against SARS-CoV-2. The C t values, age, gender and symptoms of the patients were correlated with the development of antibodies. RESULTS: IgG positivity was found to be 80.2 % (95 % CI, 76.2-84.2). Positivity increased with a decrease in the C t value, with the highest (87.6 %) positivity observed in individuals with C t values <20. The mean (±sd) C t values for IgG positives and negatives were 23.34 (±6.09) and 26.72 (±7.031), respectively. No significant difference was found for demographic characteristics such as age and sex and symptoms and antibody response. The current study is the first of its kind wherein we have assessed the correlation of the RT-PCR C t with the development of IgG against SARS-CoV-2. CONCLUSION: Although C t values might not have any relation with the development of symptoms, they are associated with the antibody response among SARS-CoV-2-infected individuals.

COVID-19 in children in Odisha state, India: a retrospective review.

We retrospectively analysed the swab samples tested for COVID-19 from 7 March 2020 to 17 August 2021 at the Indian Council of Medical Research-Regional Medical Research Centre, Bhubaneswar, Odisha. 553 763 nasopharyngeal swabs were collected from individuals suspected with COVID-19 in Odisha state. 75 190 (13.6%) samples were positive by reverse transcription-PCR. There were 5988 (8%) cases in children and young people under 18 years old. Odisha reported 996 153 COVID-19 cases which resulted in 6985 deaths in adults and 36 in children and young people under 18 years old.

Persistence of Antibodies Against Spike Glycoprotein of SARS-CoV-2 in Healthcare Workers Post Double Dose of BBV-152 and AZD1222 Vaccines.

Purpose: We investigated the persistence of the vaccine-induced immunoglobulin G (IgG) antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) among healthcare workers (HCWs) in Odisha who received a complete dose of either Covaxin or Covishield vaccine. Methods: This 24-week longitudinal cohort study was conducted from January to July 2021 with participants from 6 healthcare and research facilities of Odisha to understand the dynamicity of the vaccine-induced IgG antibodies against SARS-CoV-2 after the complete dose of vaccines. Results: Serum samples were collected from 614 participants during each follow-up and were tested in two chemiluminescent microparticle immunoassay (CLIA)-based platforms to detect SARS-CoV-2 antibodies both qualitatively and quantitatively. Among these participants, 308 (50.2%) participants were Covishield recipients and the rest 306 (49.8%) participants took Covaxin. A total of 81 breakthrough cases were recorded and the rest 533 HCWs without any history of postvaccination infection showed significant antibody waning either from T3 (Covaxin recipient) or T4 (Covishield recipient). The production of vaccine-induced IgG antibodies is significantly higher (p < 0.001) in Covishield compared with Covaxin. Covishield recipients produced higher median anti-S IgG titer than Covaxin. No statistically significant differences in antibody titers were observed based on age, gender, comorbidities, and blood groups. Conclusion: This 6-month follow-up study documents a 2-fold and 4-fold decrease in spike antibody titer among Covishield and Covaxin recipients, respectively. The clinical implications of antibody waning after vaccination are not well understood. It also highlights the need for further data to understand the long-term persistence of vaccine-induced antibody and threshold antibody titer required for protection against reinfection.