Tracking SARS-CoV-2 variants through pandemic waves using RT-PCR testing in low-resource settings.

COVID-19 resulted in extensive morbidity and mortality worldwide. SARS-CoV-2 evolved rapidly, with increasing transmission due to Variants of Concern (VOC). Identifying VOC became important but genome submissions from low-middle income countries (LMIC) remained low leading to gaps in genomic epidemi...

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Main Authors: Asghar Nasir, Uzma Bashir Aamir, Akbar Kanji, Ali Raza Bukhari, Zeeshan Ansar, Najia Karim Ghanchi, Kiran Iqbal Masood, Azra Samreen, Nazneen Islam, Samina Ghani, M Asif Syed, Mansoor Wassan, Syed Faisal Mahmood, Zahra Hasan
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2023-01-01
Series:PLOS Global Public Health
Online Access:https://doi.org/10.1371/journal.pgph.0001896
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author Asghar Nasir
Uzma Bashir Aamir
Akbar Kanji
Ali Raza Bukhari
Zeeshan Ansar
Najia Karim Ghanchi
Kiran Iqbal Masood
Azra Samreen
Nazneen Islam
Samina Ghani
M Asif Syed
Mansoor Wassan
Syed Faisal Mahmood
Zahra Hasan
author_facet Asghar Nasir
Uzma Bashir Aamir
Akbar Kanji
Ali Raza Bukhari
Zeeshan Ansar
Najia Karim Ghanchi
Kiran Iqbal Masood
Azra Samreen
Nazneen Islam
Samina Ghani
M Asif Syed
Mansoor Wassan
Syed Faisal Mahmood
Zahra Hasan
author_sort Asghar Nasir
collection DOAJ
description COVID-19 resulted in extensive morbidity and mortality worldwide. SARS-CoV-2 evolved rapidly, with increasing transmission due to Variants of Concern (VOC). Identifying VOC became important but genome submissions from low-middle income countries (LMIC) remained low leading to gaps in genomic epidemiology. We demonstrate the use of a specific mutation RT-PCR based approach to identify VOC in SARS-CoV-2 positive samples through the pandemic in Pakistan. We selected 2150 SARS-CoV-2 PCR positive respiratory specimens tested between April 2021 and February 2022, at the Aga Khan University Hospital Clinical Laboratories, Karachi, Pakistan. Commercially available RT-PCR assays were used as required for mutations in Spike protein (N501Y, A570D, E484K, K417N, L452R, P681R and deletion69_70) to identify Alpha, Beta, Gamma, Delta, and Omicron variants respectively. Three pandemic waves associated with Alpha, Delta and Omicron occurred during the study period. Of the samples screened, VOC were identified in 81.7% of cases comprising mainly; Delta (37.2%), Alpha (29.8%) and Omicron (17.1%) variants. During 2021, Alpha variants were predominant in April and May; Beta and Gamma variants emerged in May and peaked in June; the Delta variant peaked in July and remained predominant until November. Omicron (BA.1) emerged in December 2021 and remained predominant until February 2022. The CT values of Alpha, Beta, Gamma and Delta were all significantly higher than that of Omicron variants (p<0.0001). We observed VOC through the pandemic waves using spike mutation specific RT-PCR assays. We show the spike mutation specific RT-PCR assay is a rapid, low-cost and adaptable for the identification of VOC as an adjunct approach to NGS to effectively inform the public health response. Further, by associating the VOC with CT values of its diagnostic PCR we gain information regarding the viral load of samples and therefore the level of transmission and disease severity in the population.
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spelling doaj-art-103696218eee404f9190e4377c1be86d2025-08-20T02:23:19ZengPublic Library of Science (PLoS)PLOS Global Public Health2767-33752023-01-0136e000189610.1371/journal.pgph.0001896Tracking SARS-CoV-2 variants through pandemic waves using RT-PCR testing in low-resource settings.Asghar NasirUzma Bashir AamirAkbar KanjiAli Raza BukhariZeeshan AnsarNajia Karim GhanchiKiran Iqbal MasoodAzra SamreenNazneen IslamSamina GhaniM Asif SyedMansoor WassanSyed Faisal MahmoodZahra HasanCOVID-19 resulted in extensive morbidity and mortality worldwide. SARS-CoV-2 evolved rapidly, with increasing transmission due to Variants of Concern (VOC). Identifying VOC became important but genome submissions from low-middle income countries (LMIC) remained low leading to gaps in genomic epidemiology. We demonstrate the use of a specific mutation RT-PCR based approach to identify VOC in SARS-CoV-2 positive samples through the pandemic in Pakistan. We selected 2150 SARS-CoV-2 PCR positive respiratory specimens tested between April 2021 and February 2022, at the Aga Khan University Hospital Clinical Laboratories, Karachi, Pakistan. Commercially available RT-PCR assays were used as required for mutations in Spike protein (N501Y, A570D, E484K, K417N, L452R, P681R and deletion69_70) to identify Alpha, Beta, Gamma, Delta, and Omicron variants respectively. Three pandemic waves associated with Alpha, Delta and Omicron occurred during the study period. Of the samples screened, VOC were identified in 81.7% of cases comprising mainly; Delta (37.2%), Alpha (29.8%) and Omicron (17.1%) variants. During 2021, Alpha variants were predominant in April and May; Beta and Gamma variants emerged in May and peaked in June; the Delta variant peaked in July and remained predominant until November. Omicron (BA.1) emerged in December 2021 and remained predominant until February 2022. The CT values of Alpha, Beta, Gamma and Delta were all significantly higher than that of Omicron variants (p<0.0001). We observed VOC through the pandemic waves using spike mutation specific RT-PCR assays. We show the spike mutation specific RT-PCR assay is a rapid, low-cost and adaptable for the identification of VOC as an adjunct approach to NGS to effectively inform the public health response. Further, by associating the VOC with CT values of its diagnostic PCR we gain information regarding the viral load of samples and therefore the level of transmission and disease severity in the population.https://doi.org/10.1371/journal.pgph.0001896
spellingShingle Asghar Nasir
Uzma Bashir Aamir
Akbar Kanji
Ali Raza Bukhari
Zeeshan Ansar
Najia Karim Ghanchi
Kiran Iqbal Masood
Azra Samreen
Nazneen Islam
Samina Ghani
M Asif Syed
Mansoor Wassan
Syed Faisal Mahmood
Zahra Hasan
Tracking SARS-CoV-2 variants through pandemic waves using RT-PCR testing in low-resource settings.
PLOS Global Public Health
title Tracking SARS-CoV-2 variants through pandemic waves using RT-PCR testing in low-resource settings.
title_full Tracking SARS-CoV-2 variants through pandemic waves using RT-PCR testing in low-resource settings.
title_fullStr Tracking SARS-CoV-2 variants through pandemic waves using RT-PCR testing in low-resource settings.
title_full_unstemmed Tracking SARS-CoV-2 variants through pandemic waves using RT-PCR testing in low-resource settings.
title_short Tracking SARS-CoV-2 variants through pandemic waves using RT-PCR testing in low-resource settings.
title_sort tracking sars cov 2 variants through pandemic waves using rt pcr testing in low resource settings
url https://doi.org/10.1371/journal.pgph.0001896
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