Coronavirus Disease 2019 (COVID-19) caused by SARS-CoV-2 poses a significant threat to global public health. Early detection with reliable, fast, and simple assays is crucial to contain the spread of SARS-CoV-2. The real-time reverse transcription-polymerase chain reaction (RT-PCR) assay is currently the gold standard for SARS-CoV-2 detection; however, the reverse transcription loop-mediated isothermal amplification method (RT-LAMP) assay may allow for faster, simpler and cheaper screening of SARS-CoV-2. In this study, the triple-target RT-LAMP assay was first established to simultaneously detect three different target regions (ORF1ab, N and E genes) of SARS-CoV-2. The results revealed that the developed triplex RT-LAMP assay was able to detect down to 11 copies of SARS-CoV-2 RNA per 25 µL reaction, with greater sensitivity than singleplex or duplex RT-LAMP assays. Moreover, two different indicators, hydroxy naphthol blue (HNB) and cresol red, were studied in the colorimetric RT-LAMP assay; our results suggest that both indicators are suitable for RT-LAMP reactions with an obvious color change. In conclusion, our developed triplex colorimetric RT-LAMP assay may be useful for the screening of COVID-19 cases in limited-resource areas.
Background: COVID-19, caused by SARS-CoV-2, has spread worldwide and become a global pandemic. Objectives: Studies of the earliest events of the COVID-19 pandemic are critical in understanding how the pandemic started and providing insight into the spread of SARS-CoV-2 and its infection in humans. Methods: In this report, we studied the epidemiological characteristics of all 34 confirmed COVID-19 cases in Wuhu, China, from January 3 to March 19, 2020. Results: Our study indicated that cases in male patients (61.76%, 21/34) outnumbered those in female patients (38.24%, 13/34). Studies of the age distribution among the confirmed cases revealed that most COVID-19 patients were 15 to 59 years of age (26/34, 76%), while more than 14% (5/34) were ≥ 60 years old, and less than 9% (3/34) were ≤ 14 years old. Notably, 32 of the 34 confirmed cases were (a) people who had recently resided in or traveled to Wuhan or had close contact with Wuhan residents or visitors (22 cases); and (b) people who had close contact with these 22 confirmed COVID-19 patients (10 cases). Conclusions: This study revealed the epidemiological characteristics of COVID-19 outbreaks in Wuhu between January and March 2020 and provided insight into the earliest events of the COVID-19 pandemic in China. Our analyses suggested that the COVID-19 cases confirmed in Wuhu in 2020 were directly related to or originated from the outbreaks in Wuhan.
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