Introduction: Herpes simplex virus encephalitis (HSE) is an acute infection accompanied by significant morbidity and mortality with the diagnosis often made by cerebrospinal fluid (CSF) polymerase chain reaction (PCR) testing. Case Presentation: We report a case of a healthy 35-year-old woman presenting with altered mental status. Due to suspicion of herpes encephalitis, a CSF PCR for herpes virus was sent for examination and acyclovir was started. The patient had an immediate response to acyclovir; however, when the PCR returned negative she was discharged without therapy. The altered mental status returned and she was started on acyclovir therapy and a second CSF PCR sample was sent and was again negative. MRI performed at initial hospitalization was negative, but a repeat MRI demonstrated bilateral temporal lobe involvement suggestive of herpes encephalitis. The patient was successfully treated for 21 days with acyclovir. Conclusion: CSF PCR for herpes virus is highly sensitive and specific and remains the standard for diagnosing herpes encephalitis. Clinicians should be aware of the pitfalls of CSF PCR testing, specifically false-negative results. Although rare, these false negatives can result in premature termination of treatment.