Evaluating Saliva Sampling with Reverse Transcription Loop-mediated Isothermal Amplification to Improve Access to SARS-CoV-2 Diagnosis in Low-Resource Settings

Suwarti Suwarti, Sabighoh Zanjabila, Bonifacius, Yacobus Da Costa, Claus Bogh, Decy Subekti, Jeny Jeny, Ayu Madri Dewi, Nunung Nuraeni, Mutia Rahardjani, Iqbal Elyazar, Erni J. Nelwan, Anuraj H. Shankar, J. Kevin Baird, Raph L. Hamers

Research output: Contribution to journalArticlepeer-review

Abstract

Standard diagnosis of SARS-CoV-2 by nasopharyngeal swab (NPS) and real-time reverse transcriptase-polymerase chain reaction (PCR) requires a sophisticated laboratory, skilled staff, and expensive reagents that are difficult to establish and maintain in isolated, low-resource settings. In the remote setting of tropical Sumba Island, eastern Indonesia, we evaluated alternative sampling with fresh saliva (FS) and testing with colorimetric loop-medicated isothermal amplification (LAMP). Between August 2020 and May 2021, we enrolled 159 patients with suspected SARS-CoV-2 infection, of whom 75 (47%) had a positive PCR on NPS (median cycle threshold [Ct] value: 27.6, interquartile range: 12.5–37.6). PCR on FS had a sensitivity of 72.5% (50/69, 95% confidence interval [CI]: 60.4–82.5) and a specificity of 85.7% (66/77, 95% CI: 75.9–92.6), and positive (PPV) and negative (NPV) predictive values of 82.0% (95% CI: 0.0–90.6) and 77.6% (95% CI: 67.3–86.0), respectively. LAMP on NPS had a sensitivity of 68.0% (51/75, 95% CI: 56.2–78.3) and a specificity of 70.8% (63/84, 95% CI: 58.9–81.0), with PPV 70.8% (95% CI: 58.9-81.0) and NPV 72.4% (95% CI: 61.8–81.5%). LAMP on FS had a sensitivity of 62.3% (43/69, 95% CI: 49.8–73.7%) and a specificity of 72.7% (56/77, 95% CI: 61.4–82.3%), with PPV 67.2% (95% CI: 54.3–78.4) and NPV 68.3% (95% CI: 57.1–78.1%). LAMP sensitivity was higher for NPS and FS specimens with high viral loads (87.1% and 75.0% for Ct value, 26, respectively). Dried saliva on filter paper was stable for 4 days at room temperature. LAMP on either NPS or FS could offer an accessible alternative for SARS-CoV-2 diagnosis in low-resource settings, with potential for optimizing sample collection and processing, and selection of gene targets.

Original languageEnglish
Pages (from-to)284-290
Number of pages7
JournalAmerican Journal of Tropical Medicine and Hygiene
Volume107
Issue number2
DOIs
Publication statusPublished - Aug 2022

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