Inspectors have raised "serious" concerns about the cleanliness of the Accident and Emergency (A&E) ward at Kirkcaldy's Victoria Hospital.
An inspection carried out in December found dusty equipment as well as trolleys and chairs contaminated by blood and body fluids.
Concerns were also raised about dirty equipment in the intensive care unit, including intravenous pumps and syringe drivers "visibly contaminated with blood, medication, dust and enteral feed".
Inspectors found some staff were not following the proper procedures for hand washing and discovered a "significant number" of dirty mattresses.
Susan Brimelow, chief inspector of the Healthcare Environment Inspectorate (HEI), which was set up to help reduce the infection risk to patients, said she was "extremely disappointed" and had escalated her concerns to the Scottish Government.
She said: "During our inspection on December 2-3, we found patient equipment which was not clean and a poor standard of environmental cleanliness. We also found staff did not always wash their hands at appropriate times.
"We escalated our concerns to senior management and requested that immediate action be taken to minimise the risk of infection.
"On December 11, we returned unannounced to ensure improvements had been made and to assess progress. Although we saw some areas of improvement, issues with the cleanliness of patient equipment and the environment remained - in particular the A&E department and ward 15.
"Due to the serious nature of our findings, I escalated my concerns to Scottish Government."
HEI has identified seven requirements and two recommendations for NHS Fife, including ensuring that all patient equipment is clean and that all staff decontaminate their hands properly.
The health board has also been asked to investigate why cleaning standards in A&E were poor and to take action to address the problem.
Ms Brimelow added: "We will continue to inspect Victoria Hospital to ensure that improvements are made, as every patient coming into hospital should expect their local hospital to be clean."
Inspectors noted that patients who were spoken to during the visits complimented the care and treatment they received and made positive comments about ward cleanliness.
The health board's chairman Allan Burns said: "NHS Fife is constantly striving to raise the standard of care it provides and therefore it is extremely disappointing to receive this report.
"People served by the board have my assurance that everything will be done to address the concerns highlighted in the report and to attain the standards required."
Interim chief executive Dr Brian Montgomery added: "While there has been some progress in relation to areas for improvement highlighted in previous reports, there is still work to be done to raise standards to a consistent level in all areas of the Victoria Hospital.
"I would reassure our patients that urgent action has already been taken to address the issues raised by the inspection report.
"Actions already taken include the revision and enhancement of cleaning schedules, with additional monitoring at both ward and hospital management level to ensure that high standards of cleanliness are met in all wards and departments.
"Progress is being closely monitored and will be reported back directly through the NHS Fife Board. In addition, we have invited senior staff from Health Protection Scotland to provide independent scrutiny of our improvements.
"NHS Fife is fully committed to providing a clean, safe environment."
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