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.

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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

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Published

2024-10-01

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 Oct. 1 [cited 2024 Dec. 22];68(4). Available from: https://he05.tci-thaijo.org/index.php/CMJ/article/view/3261