Herpes Simplex Virus (HSV) 1 & 2, DNA Detection and Genotype, Blood or Other Specimens  

IMPORTANT! Pre-approval needed from state or local health department. Local or State Health Department. Send approval to NPHL@unmc.edu or call (402) 559-9444.

NPHL has moved to an electronic ordering system, NUlirt, and it is now the preferred method to order all tests. Once ordered, print a "batch list" (see NUlirt Guide).

Forms Required - Nulirt batch list or NPHL Test Request Form (completed in entirety) must accompany the specimen to include the following information: symptoms and date of onset, pregnancy status, travel and vaccine history. Testing will not be performed without all required information.

Please call NPHL Client Services at (402) 559-2440 or toll free at (800) 334-0459 for further clarification.


Test: Herpes Simplex Virus (HSV) 1 & 2, DNA Detection and Genotype, Blood or Other Specimens
Order in NUlirt or complete NPHL Test Order Form   
Synonym: HAA; HBsAg; Hepatitis Associated Antigen  
Method: Real-time DNA Amplification.  
Availability: Monday - Friday, Results in 24 hours.  
Specimen: Blood  
Collection Device: Adult Collection:
One 5ml Plasma Preparation tube (PPT, White) tube. One 5ml EDTA (Lavender) tube is also acceptable. Collect specimen and process using sterile technique.  For PPT tubes, plasma spin within 6h of collection and then refrigerate. 

For EDTA lavender tubes, take plasma off cells within 48h and then refrigerate. Call Molecular Virology at 402-559-7744 with any questions.
Bone marrow in EDTA, bronchial wash/bronchoalveolar lavage (BAL)/sputum, cerebral spinal fluid (CSF), tissue (fixed or snap frozen), and body fluid.
Collect tissue and process using sterile technique, (sterile blades, and containers).

Pediatric Collection:
One 5mL PPT (white) tube (Minimum 2 mL)
One 5mL EDTA (Lavender) tube. (minimum 2mL)
1mL plasma. (minimum 0.5ml plasma)
Specimen Labeling: Test subject to CLIA regulations and requires 2 patient identifiers and specimen source on container.  
Volume: Adult collection: 1mL plasma (0.5 mL minimum)  

Prior to and during shipment:  Refrigerate 2-8°C. If sample is not transported to lab the same day as collected; aliquot the serum or plasma. To aliquot: Centrifuge the sample and transfer into a clean tube and label as "serum" or "plasma."
Shipping instructions, including specimen-handling requirements during transport:  Seal screw-top tubes with Parafilm™ or tape and place in biohazard bag with absorbent material.
Place specimen inside leak proof secondary container (biohazard bag and if transporting by air, use Tyvek™ Envelope) with adsorbent material. The secondary container must be packed inside an approved Category B shipping container filled with frozen gel-packs to keep specimen refrigerated. Position NUlirt packing list or NPHL Test Order form between secondary container and box lid. Seal with tape.  Label outer box as UN3373 Category B, Biological Substance. Transport by NPHL ground courier. If courier not available, ship by FedEx, UPS, or USPS (1st Class, Priority or Express Mail). Follow appropriate DOT/IATA Regulation Guidelines.  For assistance, call NPHL State Training Coordinator at (402) 559-3590.  
See instructions and shipping address: http://nphl.org/

NUlirt packing list or appropriate paperwork (completed in entirety) must accompany specimens.

Unacceptable: Swabs: Calcium alginate or wooden shaft tips are not recommended as they may be inhibitory to DNA Amplification. Nasopharyngeal swabs are not acceptable. Swabs transported in Saline at ambient temperature. For Swab samples, please order HSV DNA direct detection, Swab.  
Specimen Stability: After separation from cells: Ambient: <8h; Refrigerated: 7d; For longer storage, freeze at -20º Celsius. (avoid more than 2 freeze/thaw cycles)  
Reference Interval: Negative, HSV-1 DNA not detected (<100 copy/mL)
Negative, HSV-2 DNA not detected (<500 copy/mL)  
Reportable Disease: Immediately report positive Hepatitis B results to local or state health department. Refer to Nebraska DHHS Title 173, Communicable Diseases.   
Comments: Pre-approval needed from state or local health department.  Send approval to NPHL@unmc.edu or call (402) 559-9444.

Herpes Simplex viruses (HSV) 1 and 2 are double stranded DNA viruses of the Herpesviridae family, sharing over half of the same genome. Herpes viruses may cause a wide range of diseases in neonates and immuno-compromised patients. HSV-1 typically causes orofacial lesions, keratitis, pneumonia, or encephalitis.  HSV-2 typically causes genital lesions or meningitis.  Both can cause severe neonatal herpes disease when transmitted during childbirth1.    The Real Time amplification enables the detection, differentiation, and reporting of HSV-1 and HSV-2 to be shortened, allowing for rapid turnaround time and diagnosis.

This method integrates fluorogenic DNA amplification with Real-Time PCR based detection to detect specific PCR amplicons using scorpion primers specific for HSV-1 and HSV-2. This real-time test targets the DNA polymerase gene for HSV-1 and HSV- 2. Primers are ASR, precise nucleic acid sequence information is not provided by the manufacturer.

NUlirt packing list or NPHL Test Order Form (completed in entirety) must accompany the specimen.  
Revised: 6/11/2021 
If you have questions about proper specimen collection, please call NPHL Client Services at (402) 559-2440 or toll free at (800) 334-0459.