Specialized and Forensic Collections

Prostatitis localization

The Meares-Stamey four-glass protocol is the reference localization method for suspected chronic bacterial prostatitis. The 2026 European Association of Urology (EAU) guideline identifies it as the optimum test and also recommends the two-glass pre- and post-massage test. The two-glass test requires two specimens instead of four and has similar diagnostic sensitivity.^{11 European Association of Urology. EAU Guidelines on Urological Infections. 2026.} The four-glass protocol collects voided bladder (VB) urine portions and expressed prostatic secretion (EPS):

The laboratory cultures VB1, VB2, EPS, and VB3 quantitatively and examines the urine portions and EPS for leukocytes. A prostatic source is supported when an organism grows in EPS or VB3 and is absent from both VB1 and VB2. It is also supported when the count in EPS or VB3 exceeds the pre-massage count by the threshold specified in the laboratory procedure.

The two-glass test compares a clean-catch urine specimen collected before prostatic massage with the first urine passed after massage. These specimens correspond to VB2 and VB3, respectively. A 353-participant comparison found strong concordance with the four-glass protocol, and the 2026 EAU guideline reports similar diagnostic sensitivity. Either method may be used for chronic bacterial prostatitis.^{1,21 European Association of Urology. EAU Guidelines on Urological Infections. 2026.2 Nickel JC, et al. How does the pre-massage and post-massage 2-glass test compare… 2006.} For primary prostate pain syndrome, the EAU describes the four-glass protocol as too complex for routine urologic use and the two-glass test as a cost-effective screening procedure. It also reports limited diagnostic yield. Positive localization cultures occurred in 8% of patients with suspected primary prostate pain syndrome, a rate similar to that in asymptomatic males. These cultures therefore do not establish the pain syndrome and are not a general test for chronic pelvic pain.^{33 European Association of Urology. EAU Guidelines on Chronic Pelvic Pain. 2026.}

EAU guidance and the Nickel et al. comparison use different culture-count ratios and leukocyte cutoffs. The laboratory procedure must state the criteria used to identify prostatic localization and inflammation, so every result is read against the same cutoffs.^{1,21 European Association of Urology. EAU Guidelines on Urological Infections. 2026.2 Nickel JC, et al. How does the pre-massage and post-massage 2-glass test compare… 2006.}

Federal workplace drug testing

Federal workplace urine drug testing uses chain-of-custody documentation from collection through testing. The 2026 Federal Custody and Control Form (Federal CCF) records each custody transfer and the condition of the primary specimen seal when the test facility receives the package.^{4,54 Substance Abuse and Mental Health Services… Urine Specimen Collection Handbook for Federal Agency Workplace… 2024.5 Substance Abuse and Mental Health Services… Guidance for Using the 2026 Federal Custody and Control Form… 2026.} The Substance Abuse and Mental Health Services Administration permits continued use of the 2020 Federal CCF and its September 2023 revision without a memorandum for record until May 31, 2027. As of that date, the test facility treats use of an older form as a correctable discrepancy under the current guidance. The collector and test facility follow the form and transition instructions in effect on the collection date.^{55 Substance Abuse and Mental Health Services… Guidance for Using the 2026 Federal Custody and Control Form… 2026.}

For US federal workplace urine testing, the following requirements apply.^{4,54 Substance Abuse and Mental Health Services… Urine Specimen Collection Handbook for Federal Agency Workplace… 2024.5 Substance Abuse and Mental Health Services… Guidance for Using the 2026 Federal Custody and Control Form… 2026.}

• The donor provides at least 45 mL of urine. The collector places at least 30 mL in primary bottle A and at least 15 mL in split bottle B.
• The collector measures specimen temperature within 4 minutes after receiving the specimen. The acceptable range is 32 to 38 °C (90 to 100 °F). A temperature outside this range requires a second collection under direct observation.
• The donor watches the collector seal both bottles, then initials the tamper-evident seals and signs the donor certification on the Federal CCF. The collector completes the remaining required entries and sends the sealed package within 24 hours to a laboratory or instrumented initial test facility (IITF) certified by the Department of Health and Human Services (HHS).
• If the 24-hour window cannot be met, the collector sends the sealed package on the next business day.

Whenever shipment will exceed 24 hours, the collector keeps the packaged specimens in secure refrigerated storage until shipment, as directed by the federal collection handbook.^{44 Substance Abuse and Mental Health Services… Urine Specimen Collection Handbook for Federal Agency Workplace… 2024.}

At receipt, the HHS-certified laboratory or IITF continues the chain of custody in Step 4, records the receipt date and primary seal condition, then records release to storage or another controlled location. When an IITF forwards a urine specimen to a laboratory, the laboratory continues the chain on the IITF Supplemental CCF.^{55 Substance Abuse and Mental Health Services… Guidance for Using the 2026 Federal Custody and Control Form… 2026.}

The HHS-certified laboratory classifies a specimen as dilute, substituted, adulterated, or invalid. Classification follows the creatinine, specific-gravity, pH, and confirmatory-test criteria in the federal Mandatory Guidelines. Nonfederal programs follow their own requirements and may use different collection volumes, temperature limits, or procedures.^{66 Mandatory Guidelines for Federal Workplace…, et al. 88 Fed Reg 70768-70810. 2023.} Confirm which program governs a specimen before you handle it, because the program sets the volumes, limits, and custody steps.

Practice