After applying Windows Updates to a PVS vdisk using a master target, and then later booting it on production PVS targets, a boot time error "LoadImage error: Access Denied" is displayed.
Only the master target, which was used when applying the windows update continues to boot.

The vDisk has been updated, so that the Windows Boot Loader is signed by a new certificate.
As documented by Microsoft (see additional information), the current certificates used for validating binaries when using Secure Boot are expiring in 2026 and have been replaced by new CA certificates which will be used in the future to validate that binaries are properly signed. When applying Windows Updates, if the new Windows certificate is detected in the master VM, it will install a version of the Boot Loader that is signed by the new certificate. Later on, if you have target VMs that do not include the new certificate, the resulting vDIsk will not be bootable as it cannot validate the signature of the Boot Loader.
When you upgrade your hypervisor to a version that embeds the new certificates, existing VMs are not updated to include this new certificate by default which is why subsequent boot of the vDisk fails with the above error.
NOTE: the expiration of the existing certificates does not matter and existing vDisks will continue to boot after expiration. The only failure occurs when a new Windows Boot Loader is installed in the vDisk that is signed by the new certificate.
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This behavior would follow a pattern like below:
The following steps should be followed for VMWare and XenServer deployments:
Important Notes
This article shows how to address the issue for VMWare and XenServer based setups, where PVS Targets using Secure Boot Become Unbootable After Applying Windows Updates to the vDisk
Windows Secure Boot certificate expiration and CA updates
https://support.microsoft.com/en-us/topic/windows-secure-boot-certificate-expiration-and-ca-updates-7ff40d33-95dc-4c3c-8725-a9b95457578e