Retrospective analysis of SARS-CoV-2 RNA detection in pooled saliva samples: An effective cost-saving method

Authors

  • Jiraphat Charoenkupt King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand
  • Ati Burassakarn Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
  • Arkom Chaiwongkot Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

Keywords:

COVID-19, pooling saliva samples, RT-PCR, SARS-CoV-2

Abstract

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a causative agent of coronavirus disease 2019 (COVID-19), which was first reported in December 2019 and has since spread globally. Effective laboratory testing is crucial for the early detection and prevention of SARS-CoV-2 trans mission. The use of pooled saliva samples represents a potential method for active case finding and increasing testing efficiency.

Objectives: To evaluate the effectiveness of pooled saliva samples for SARS-CoV-2 testing and its costeffectiveness for screening healthcare workers at King Chulalongkorn Memorial Hospital, Thai Red Cross Society.

Methods: A total of 24,098 samples collected between April 19, 2021, and May 30, 2022, to be tested for the presence of SARS-CoV-2 were analyzed retrospectively. The samples were examined individually and in pools of four and six using the Cobas 6,800 reverse-transcription polymerase chain reaction assay for the detection of SARS-CoV-2 RNAs. The analysis focused on changes in cycle threshold values for each target between positive pools and positive individual samples.

Results: SARS-CoV-2 was detected in 0.5% of the samples (123/24,098). Pooling saliva samples in groups of four or six did not compromise the detection of viral RNAs. Pooled saliva testing showed high performance for SARS-CoV-2 detection, with cost reductions of 73.5% for the four-sample pools and 80.7% for the sixsample pools compared with individual testing.

Conclusion: Pooling saliva samples is a cost-effective and efficient method for screening SARS-CoV-2, particularly in low-prevalence settings. This approach helps quickly identify and isolate healthcare workers with infection, thus reducing transmission and preserving resources.

Downloads

Download data is not yet available.

References

Sarker R, Roknuzzaman ASM, Nazmunnahar, Hossain MJ, Islam MR. Benefits and probable ill effects of WHO's declaration of end of COVID-19 pandemic: a way back to pandemic-free normal life. Ann Med Surg (Lond) 2023;85:3199-201.

https://doi.org/10.1097/MS9.0000000000000848

Jacobs CM. Leadership-followership relationship through the pandemic for our first lines of defense: Healthcare workers and military personnel. Int J Nurs Health Care Res 2023;6: 1430.

https://doi.org/10.29011/2688-9501.101430

Otshudiema JO, Folefack GLT, Nsio JM, Kakema CH, Minikulu L, Bafuana A, et al. Community-based COVID-19 active case finding and rapid response in the Democratic Republic of the Congo: Improving case detection and response. PLoS One 2023;18:e0278251.

https://doi.org/10.1371/journal.pone.0278251

Procop GW, Shrestha NK, Vogel S, Van Sickle K, Harrington S, Rhoads DD,et al. A direct comparison of enhanced saliva to nasopharyngeal swab for the detection of SARS-CoV-2 in symptomatic patients. J Clin Microbiol 2020;58:e01946-20.

https://doi.org/10.1128/JCM.01946-20

Rivas-Macho A, Sorarrain A, Marimón JM, Goñi-deCerio F, Olabarria G. Extraction-free colorimetric RTLAMP detection of SARS-CoV-2 in saliva. Diagnostics (Basel) 2023;13:2344.

https://doi.org/10.3390/diagnostics13142344

Snipaitiene K, Zablockiene B, Sabaliauskaite R, Zukauskaite K, Matulyte E, Smalinskaite T, et al. SARSCoV-2 RT-qPCR Ct values in saliva and nasopharyngeal swab samples for disease severity prediction. J Oral Microbiol 2023;15:2213106.

https://doi.org/10.1080/20002297.2023.2213106

Bordi L, Sberna G, Lalle E, Fabeni L, Mazzotta V, Lanini S, et al. Comparison of SARS-CoV-2 detection in nasopharyngeal swab and saliva samples from patients infected with Omicron variant. Int J Mol Sci 2023;24:4847.

https://doi.org/10.3390/ijms24054847

Sahajpal NS, Mondal AK, Ananth S, Njau A, Ahluwalia P, Newnam G, et al. SalivaSTAT: Direct-PCR and Pooling of saliva samples collected in healthcare and community setting for SARS-CoV-2 mass surveillance. Diagnostics (Basel) 2021;11:904.

https://doi.org/10.3390/diagnostics11050904

Allicock OM, Yolda-Carr D, Todd JA, Wyllie AL. Pooled RNA-extraction-free testing of saliva for the detection of SARS-CoV-2. Sci Rep 2023;13:7426.

https://doi.org/10.1038/s41598-023-34662-2

Watkins AE, Fenichel EP, Weinberger DM, Vogels CBF, Brackney DE, Casanovas-Massana A, et al. Pooling saliva to increase SARS-CoV-2 testing capacity. medRxiv. 2020 :2020.09.02.20183830.

https://doi.org/10.1101/2020.09.02.20183830

Mahmoud S, Ganesan S, Raheja P, Cantarutti F, Ateia H, Zaher WA. Diagnostic accuracy of the Cobas 6800 RT-PCR assay for detection of SARS-CoV-2 RNA. Front Anal Sci 2022;2 :1030701.

https://doi.org/10.3389/frans.2022.1030701

Barat B, Das S, De Giorgi V, Henderson DK, Kopka S, Lau AF, et al. Pooled Saliva Specimens for SARS-CoV-2 Testing. J Clin Microbiol 2021;59:e02486-20.

https://doi.org/10.1128/JCM.02486-20

McMillen T, Jani K, Babady NE. Evaluation of sample pooling for SARS-CoV-2 RNA detection in nasopharyngeal swabs and salivas on the Roche Cobas 6800. J Clin Virol 2021;138:104790.

https://doi.org/10.1016/j.jcv.2021.104790

Mahmoud SA, Ibrahim E, Thakre B, Teddy JG, Raheja P, Ganesan S, et al. Evaluation of pooling of samples for testing SARS-CoV- 2 for mass screening of COVID-19. BMC Infect Dis 2021;21:360.

https://doi.org/10.1186/s12879-021-06061-3

Toppings NB, Oberding LK, Lin YC, Evans D, Pillai DR. The Role of Subgenomic RNA in Discordant Results From Reverse Transcription-Polymerase Chain Reaction Tests for COVID-19. Arch Pathol Lab Med 2022;146:805-13.

https://doi.org/10.5858/arpa.2021-0630-SA

Rodino KG, Peaper DR, Kelly BJ, Bushman F, Marques A, Adhikari H, et al. Partial ORF1ab Gene Target Failure with Omicron BA.2.12.1. medRxiv 2022:2022.04. 25.22274187.

https://doi.org/10.1101/2022.04.25.22274187

Derin D, Gültekin E, Taþkýn I, Otlu B, Öktem HA. Designing of rapid assay for the detection of RdRp/ Orf1ab specific to SARS-CoV-2. J Virol Methods 2023;320:114774.

https://doi.org/10.1016/j.jviromet.2023.114774

Popping S, Molenkamp R, Weigel JD, Mutsaers P, Voermans JC, van Boheemen S, et al. Diminished amplification of SARS-CoV-2 ORF1ab in a commercial dual-target qRT-PCR diagnostic assay. J Virol Methods 2022;300:114397.

https://doi.org/10.1016/j.jviromet.2021.114397

Sirijatuphat R, Leelarasamee A, Horthongkham N. Prevalence and factors associated with COVID-19 among healthcare workers at a university hospital in Thailand. Medicine (Baltimore) 2022;101:e30837.

https://doi.org/10.1097/MD.0000000000030837

Zhuang X, Zhao Z, Feng X, Lui GCY, Chan D, Lee SS, et al. Integrating Magnetic-Bead-Based Sample Extraction and Molecular Barcoding for the One-Step Pooled RT-qPCR Assay of Viral Pathogens without Retesting. Anal Chem 2023;95:6182-90.

https://doi.org/10.1021/acs.analchem.3c00885

Cabrera A, Al Mutawah F, Kadour M, Schofield S, Conkey B, Fuller J, et al. Increasing SARS-CoV-2 testing capacity through specimen pooling: An acute care center experience. PLoS One 2023;18:e0267137.

https://doi.org/10.1371/journal.pone.0267137

Wagner K, Fox P, Gordon E, Hahn W, Olsen K, Markham A, et al. A multiplexed, paired-pooled droplet digital PCR assay for detection of SARS-CoV-2 in saliva. Sci Rep 2023;13:3075.

https://doi.org/10.1038/s41598-023-29858-5

Law S, Tovar MA, Franke MF, Calderon R, Palomino S, Valderrama G, et al. Low detection rate of RT-PCRconfirmed COVID-19 using IgM/IgG rapid antibody tests in a large community sample in Lima, Peru. BMC Infect Dis 2023;23:62.

https://doi.org/10.1186/s12879-023-08003-7

Moreno-Contreras J, Espinoza MA, Sandoval-Jaime C, Cantú-Cuevas MA, Madrid-González DA, BarónOlivares H, et al. Pooling saliva samples as an excellent option to increase the surveillance for SARS-CoV-2 when re-opening community settings. PLoS One 2022;17:e0263114.

https://doi.org/10.1371/journal.pone.0263114

Downloads

Published

2024-09-27

How to Cite

1.
Charoenkupt J, Burassakarn A, Chaiwongkot A. Retrospective analysis of SARS-CoV-2 RNA detection in pooled saliva samples: An effective cost-saving method. Chula Med J [Internet]. 2024 Sep. 27 [cited 2024 Oct. 31];68(4). Available from: https://he05.tci-thaijo.org/index.php/CMJ/article/view/3261