RealLine Treponema pallidum

Clinical information:

Treponema pallidum is a motile spirochaete that is generally acquired by close sexual contact, entering the host via breaches in squamous or columnar epithelium. T. pallidum is causing Syphilis is a sexually transmitted infection. The primary route of transmission is through sexual contact; it may also be transmitted from mother to fetus during pregnancy or at birth, resulting in congenital syphilis.

Typical for this disease is a beginning with painless mucosal ulcers and swollen lymph nodes. In some of the infected there is a chronic course, which is characterized by a variety of skin and organ involvement. In the final stage it comes to the destruction of the central nervous system.

The extraction of DNA from clinical materials can be performed using the extraction kits:

• RealLine DNA-Express (REF VBC8899-R)
• RealLine Extraction 100 (REF VBC8896-R)
• INSTANT Virus RNA/DNA Kit – IPC16 (Roboscreen GmbH, Germany)
• INSTANT Virus RNA/DNA Kit – FX (Roboscreen GmbH, Germany)
• VeriLab UMag (Laboveritas SIA)

RealLine Treponema pallidum(Str-format) kit is designed for the analysis of clinical materials such as swabs of the epithelial cells, tissue fluid of erosive–ulcerative skin lesion and mucosa.

The assay is based on real-time polymerase chain reaction (PCR) method with fluorescent detection of amplified product.

The Str-Format Kit contains 48 tubes (6 x 8, 0.2 ml) in strips with lyophilized Mastermix. 50 μl of extracted DNA have to be pipetted into the tube and the ready Mastermix is diluted. The kit contains reagents required for 48 tests, including control samples.

The kit is validated for use with block-type cyclers: iQ™5 iCycler, CFX™96 (Bio-Rad, USA), DT-96 (DNA-Technology, Russia) and RealLine Cycler (BIORON Diagnostics GmbH).

The use of:

!  Extraction Kits for nucleic acids from clinical specimen from other supplier

!  other real-time PCR devices

!  appropriate reaction volumes, other than 50 µl

has to be validated in the lab by the user. The special notes regarding the internal control IC have to be strongly followed.

CE IVD

RealLine DNA-Express (REF VBC8899-R)

RealLine Extraction 100 (REF VBC8896-R)

INSTANT Virus RNA/DNA Kit – IPC16 (Roboscreen GmbH, Germany)

INSTANT Virus RNA/DNA Kit – FX (Roboscreen GmbH, Germany)

VeriLab UMag (Laboveritas SIA)

The kit is validated for use with block-type cyclers: iQ™5 iCycler, CFX™96 (Bio-Rad, USA), DT-96 (DNA-Technology, Russia) and RealLine Cycler (BIORON Diagnostics GmbH).

Sensitivity

Sensitivity control was performed on five samples containing 100 copies of Treponema pallidum DNA per sample, prepared from Standard Reference Sample (Treponema pallidum DNA SRS) as percentage of samples defined as positive.

The sensitivity equals 100%.

Specificity

Specificity of detection of Treponema pallidum DNA was confirmed in laboratory tests using negative samples of the Standard Reference Panel, as percentage of samples defined as negative.

Specificity of detection equals 100 %.

Diagnostic sensitivity

Clinical studies conducted in two independent laboratories on 44 positive samples of biological material (scrapings of epithelial cells) obtained from patients with syphilitic erosive and ulcerative lesions of the mucous membranes, showed sensitivity of 100 % (range 93 % -100 %, with a confidence level of 90 %).

Diagnostic specificity

Clinical studies conducted in two independent laboratories on the 44 negative samples of biological material (scrapings of epithelial cells) derived from apparently healthy individuals, patients with various STD (Chlamydia trachomatis, Ureaplasma species, Mycoplasma hominis, Trichomonas vaginalis, Neisseria gonorrhoeae, HSV type 1 and 2, HIV, but not syphilis) and from patients infected with HPV types 16, 18, 31, 33, 45, 51, showed specificity of 100 % (range 93 % -100 %, with a confidence level of 90 %).

• Instruction for use
• Molecular Diagnostic Products