Intended Use
The AMD Measles Virus (MeV) RT-qPCR Kit is a high-sensitivity, in-vitro Real-Time RT-PCR diagnostic tool for the qualitative detection of Measles virus RNA. Designed for In Vitro Diagnostic (IVD) use, it employs the TaqMan® detection method in a one-step RT-qPCR format optimized for accuracy, sensitivity, and reliability.

Overview
Measles continues to be one of the major causes of child illness and death globally. Caused by the Measles virus, this acute viral disease spreads via coughing, sneezing, close contact, or direct exposure to infected nasal or throat secretions. The virus can survive in the air or on surfaces for up to two hours, making it highly contagious. Outbreaks often lead to deadly epidemics, especially in malnourished or unvaccinated children.

Symptoms may begin with fever, cough, and a rash, but complications like diarrhea, ear infections, and pneumonia are common. Severe outcomes such as seizures, blindness, and encephalitis may also occur, although less frequently.

Test Principle
This kit detects Measles virus RNA through hydrolysis probe-based RT-qPCR. Initially, viral RNA is reverse transcribed into complementary DNA (cDNA). Amplification of the MeV N gene follows, using primers and probes tagged with fluorescent reporter dyes.

During the RT step, denatured RNA allows the synthesis of a single DNA strand (cDNA). This strand is then amplified by DNA polymerase to generate double-stranded cDNA. The included probe—labelled with a 5’ reporter dye and 3’ quencher—is hydrolyzed during amplification. As Taq polymerase progresses, it breaks the probe apart, separating the dye from the quencher, resulting in a measurable fluorescence signal. This signal increases in proportion to the presence of the target RNA.

The kit includes an internal positive control assay, verifying both the RNA extraction process and the absence of PCR inhibitors. Reactions are pre-optimized in a ready-to-use hot-start master mix to reduce non-specific amplification and maximize sensitivity.

Reagent Handling and Storage
Store the kit between -10°C and -30°C to ensure stability until the expiration date. Avoid repeated freeze-thaw cycles, which can reduce detection sensitivity. It’s advised to aliquot the master mix to prevent degradation. Keep reagents protected from light, and ensure all components are fully thawed, well-mixed, and centrifuged before use.

Instrument Compatibility
The AMD Measles RT-qPCR Kit is compatible with widely used real-time PCR systems capable of detecting FAM and HEX dyes, such as:

• Bio-Rad CFX96

• Applied Biosystems 7500 Fast

• QuantStudio 3, 5, 7

• Agilent Mx3000/3005P

• Rotor-Gene Q

• Cepheid Smartcycler

• Analytik Jena qTower

• Roche Lightcycler 480/96

Sample Collection, Storage & Transport
Ideal specimens include nasopharyngeal or throat swabs, though urine and blood can also be tested. Swabs are the preferred sample type for best sensitivity.

Samples should be stored at -20°C or -80°C if not used immediately. Avoid freeze-thaw cycles as they compromise test results. Transport samples in secure, shatterproof containers per local biohazard transport regulations. If possible, deliver by courier within 6 hours at 2–8°C to maintain sample integrity.