The change happened on Monday as shares in the online retail giant rose 3%, lifting its market value to $797m (£624.3m).
Microsoft also rose, but by less than 1%, leaving its value at $784bn (£614m).
It is the first time that Amazon – founded by Jeff Bezos in 1994 – has held the top spot and ends Microsoft’s brief return to the number one position after it surpassed Apple late in November.
Apple share prices have been under pressure amid fears that Donald Trump’s trade war with China will hold back economic growth.
Technology stocks and other industries that generate a substantial chunk of revenues outside the US are among those that have been badly affected by the market volatility.
Amazon, despite climbing ahead of Microsoft, is worth a fifth less than in September when its value peaked at more than $1tn.
Apple was worth even more at the time but its stock has plunged by 37% since mid-October, shaving $400bn (£313bn) off its valuation.
The iPhone maker confirmed some of investors’ worst fears last week when it warned it would miss its quarterly revenue target, largely blaming a slowdown in China amid its trade war with the US.
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