Speaking at the NHS Confederation conference, chief nursing officer Christine Beasley said the culture in which infection was not seen as important needed to change.
The talk coincided with the launch of the updated version of Saving Lives, which if used can lead to hygiene code compliance, says the DoH.
Ms Beasley said: 'Infection causes pain and suffering and sometimes death. Although the chances are still relatively small, whether it?s right or wrong I wouldn't mind betting that if you were going in to hospital for an operation, you would have in the back of your mind: 'I hope I don't get an infection'.
She said: 'If clinicians have one patient with MRSA they say: 'It's only one.' We need to get rid of that kind of culture. We need zero tolerance.'
Ms Beasley also commented on the new BUPA advert which states their hospitals are 'clean' and highlighted the fact that patients use cleanliness as a touchstone to judge how the organisation is performing.
The DoH maintains it will hit the target to halve instances of MRSA by April 2008, contrary to a leaked document seen by HSJin January.
Director of performance at the department Richard Gleave said: 'The target is challenging but doable - it can be done we are very clear about that.
'The target is a litmus test and we can't pretend we have made progress if we can't hit the target.'
The DoH has begun talks with the NHS Confederation on developing a 'relevant' way to publish data on MRSA currently published by the Health Protection Agency.
Ms Beasley said that healthcare-acquired infection costs at least£4,000-10,000 per patient; it can result in a 21-day stay which quickly adds up to real money and it impacts on how patients can be moved around.