Forty-six male and 248 female healthy volunteers, who were 20 to 65 years old, were recruited from the general population in Shanghai between April and May 2017. Medical and medication history was obtained for each individual by questionnaires. Subjects with any history of skin diseases and intake of systemic or local antibiotics in the past 6 months were excluded. To maximize microbial skin load, each subject was instructed to wash the face only with tap water and to refrain from the application of any skin-care or cosmetic products on the sampling day before sampling.

Three skin sites (forehead, cheek, the back of the nose) were sampled for each subject. Study personnel wore sterile gloves for each sample collection. Samples were collected in a temperature and humidity-controlled room at 20 °C and 50% humidity. To obtain sufficient DNA from the three anatomical skin sites, which were low and variable in microbial biomass, and for the sake of establishing uniform standards between samples, a skin area of 4 cm2 was swabbed by sterile polyester fiber-headed swabs moistened with a solution of 0.15 M NaCl and 0.1% Tween 20 [60]. The sampling regions were swabbed 40 times each. Then, the swab head was fractured, placed in a sterilized 1.5 mL centrifuge tube, and stored at − 80 °C [9].

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