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The BD ProbeTec™ Herpes Simplex Viruses (HSV 1 & 2) Q^x Amplified DNA Assays (HSV Q^x Assays), when tested with the BD Viper™ System in Extracted Mode, use Strand Displacement Amplification technology for the direct, qualitative detection and differentiation of Herpes Simplex virus type 1 (HSV1) and Herpes Simplex virus type 2 (HSV2) DNA in clinician-collected external anogenital lesion specimens. The assays are indicated for use with symptomatic individuals to aid in the diagnosis of anogenital HSV1 and HSV2 infections.

Herpes Simplex virus type 1 (HSV1) and type 2 (HSV2) are ubiquitous double-stranded neurotropic DNA viruses of the Herpesviridae family that cause incurable lifelong infections and which result from inoculation of the virus through abraded skin or mucous membranes. Both viruses may remain latent for long periods and cause recurrent episodes of symptomatic disease.

The primary modes of transmission for HSV1 are via oral secretions and non-genital contact, resulting in a predominance of infections of the oropharynx, face, eyes and central nervous system. In recent years the frequency of genital HSV1 infections has increased due to a lower rate of oral infection during prepubescent childhood that renders individuals susceptible to genital infection in later life and a rise in the frequency of oro-genital contact.^1,2 Nevertheless, seroprevalence studies show that up to 80% of children are infected with HSV1 by adulthood, with the highest rates among those in poor socioeconomic groups.³

In contrast with HSV1, infection with HSV2 is usually the result of sexual transmission. In the United States, 20 – 25% of the population has antibodies for HSV2 by the age of 40 and overall there are at least 50 million people with genital herpes.³ In the majority of cases, symptoms are mild or unrecognized and the infection remains undiagnosed, although intermittent shedding of infectious virus into the genital tract still occurs. As a result, most transmission of genital herpes occurs through sexual contact by persons who are asymptomatic or are unaware that they are infected. Infection with HSV2 is more common in women than men and is linked to an increased risk of sexually transmitted Human Immunodeficiency Virus (HIV).⁴ While transmission of herpes to neonates in utero or intrapartum is rare, the consequences of such infections are severe and frequently fatal.⁵

Classical primary genital herpes infection is preceded by localized pain or tingling, frequently accompanied by fever, malaise and inguinal lymphadenopathy. Within days, vesicles appear on the labia minora, introitus and urethra meatus of women and on the shaft and glans of the penis in men. The perineum and perianal areas may also be affected, as may the upper thighs and buttocks. Cervical lesions also occur frequently in women. Primary infection with HSV1 cannot be distinguished from that caused by HSV2 on clinical grounds and the lesions may also be confused with those caused by other sexually transmitted diseases. As a consequence, laboratory testing is required for definitive diagnosis to reduce symptoms and hasten the healing of lesions. In addition, because the recurrence of HSV1 infections and subclinical shedding are less frequent than for HSV2, determination of the etiology of infection and typing of the virus is useful in the assessment of prognosis and counseling.^1,2,6 The preferred method for diagnosis of herpes infection has historically been viral isolation in tissue culture followed by type-specific immunofluorescent detection; however, the enhanced sensitivity, robustness and rapid time-toresults of amplified methods for the detection of viral DNA are leading to their increasingly widespread adoption.^1,3,6

When used with the BD Viper System in extracted mode, the BD ProbeTec HSV Q^x Amplified DNA Assays involve automated non-specific extraction of DNA from clinical specimens using chemical lysis, followed by binding of DNA to magnetic particles, washing of the bound nucleic acid and elution in an amplification-compatible buffer. When present, HSV1 and/or 2 DNA is detected by Strand Displacement Amplification (SDA) of type-specific target sequences in the presence of a fluorescently labeled detector probe.^7,8

Quality control must be performed in accordance with applicable local, state and/or federal regulations or accreditation requirements and your laboratory's standard Quality Control procedures. It is recommended that the user refer to pertinent CLSI guidance and CLIA regulations for appropriate Quality Control practices.

The Control Set for the BD ProbeTec HSV Q^x Amplified DNA Assays is provided separately. One Positive and one Negative Control must be included on each plate and for each new reagent kit lot number. Controls must be positioned according to the BD Viper Instrument User's Manual. The HSV Q^x Positive Control will monitor for substantial reagent failure only. The HSV Q^x Negative Control monitors for reagent and/or environmental contamination.

The HSV Q^x Positive Control contains cloned HSV1 and HSV2 target regions. These controls may be used for internal quality control or users may develop their own internal quality control material.¹³ Additional controls may be tested according to guidelines or requirements of local, state, and/or federal regulations or accrediting organizations. Refer to CLSI C24-A3 for additional guidance on appropriate internal quality control testing practices.¹³ The Positive Control contains approximately 15,000 copies per mL of pHSV1 and 16,500 copies per mL of pHSV2 linearized plasmids.

The Extraction Control (EC) oligonucleotide is used to confirm the validity of the extraction process. The EC is dried in the Extraction Tubes and is re-hydrated by the BD Viper System upon addition of the specimen and extraction reagents. At the end of the extraction process, the EC fluorescence is monitored by the instrument and an automated algorithm is applied to both the EC and the HSV specific signals to report specimen results as positive, negative, or EC failure.

Each BD ProbeTec HSV Q^x Reagent Pack contains:

HSV1 Q^x Amplified DNA Assay Priming Microwells, 3 pouches of 32 microwells each: each Priming Microwell contains approximately 108 pmol oligonucleotides, 27 pmol fluorescently labeled detector probe, 150 nmol dNTPs, with stabilizers and buffer components.

HSV1 Q^x Amplified DNA Assay Amplification Microwells, 3 pouches of 32 microwells each: each Amplification Microwell contains approximately 100 units DNA polymerase and 500 units restriction enzyme, with stabilizers and buffer components.

HSV2 Q^x Amplified DNA Assay Priming Microwells, 3 pouches of 32 microwells each: each Priming Microwell contains approximately 105 pmol oligonucleotides, 36 pmol fluorescently labeled detector probe, 120 nmol dNTPs, with stabilizers and buffer components.

HSV2 Q^x Amplified DNA Assay Amplification Microwells, 3 pouches of 32 microwells each: each Amplification Microwell contains approximately 35 units DNA polymerase and 500 units restriction enzyme, with stabilizers and buffer components.

NOTE: Each microwell pouch contains one desiccant bag.

General

1. For in vitro Diagnostic Use.

2. Pathogenic microorganisms, including hepatitis viruses and Human Immunodeficiency Virus, may be present in clinical specimens. "Standard Precautions"^9-12 and institutional guidelines should be followed in handling all items contaminated with blood and other body fluids.

3. For additional specific warnings, cautions, and notes specific to the BD Viper, consult the BD Viper Instrument User's Manual.