ハトパラミクソウイルス1型(PPMV-1)RNA検査キット - RT-qPCR

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カタログ番号 AP2409006  Specification: 50T / 100Tストレージ: –20°C


Product Overview:

について ハトパラミクソウイルス1型(PPMV-1)RNA検出キット is a highly sensitive RT-qPCR diagnostic tool for identifying PPMV-1, an RNA virus that causes Newcastle disease in pigeons. Clinical signs in infected birds include diarrhea, leg paralysis, tremors, and neurological symptoms.

PPMV-1 is highly contagious, with a high morbidity and mortality rate. It spreads rapidly via oral, respiratory, ocular, reproductive, and even wound-related routes. Timely and accurate detection is essential for disease control, prevention of secondary infections, and containment of outbreaks.

This kit is based on conserved gene sequences of PPMV-1 and has been optimized for rapid detection in oral/cloacal swabs, blood, and tissues of pigeons and other avian species via one-step reverse transcription quantitative PCR (RT-qPCR).


Key Features:

  • One-step RT-qPCR for direct RNA detection

  • Applicable to swabs, blood (EDTA), tissue homogenates

  • Includes positive and negative controls

  • Fast results, high sensitivity, and specificity

  • Long shelf life: ≥12 months at –20°C

  • Ideal for pigeon breeders, avian laboratories, and veterinary surveillance


Kit Components:

Component50T100T
PPMV-1 Reaction Buffer575 µL1150 µL
PPMV-1 Premix575 µL1150 µL
PPMV-1 Positive Control100 µL100 µL
Negative Control100 µL100 µL

Sample Types & Requirements:

  • Oral/Cloacal Swabs: Sterile swabs stored at 2–8°C (within 24h) or –20°C

  • Whole Blood: Use only EDTA anticoagulant, avoid heparin; store ≤7 days at 2–8°C or ≤3 months at –20°C

  • Tissues: ≤100 mg of fresh muscle or organ tissue, homogenized in 200–500 µL sterile water

RNA should be extracted using the MD023 Safe RNA Extraction Kit or equivalent. Extracted RNA should be stored at 4°C (≤7 days) or –20°C (≤6 months).


RT-qPCR Reaction Setup:

  1. Thaw reagents and centrifuge briefly.

  2. For n samples + 2 controls, mix:

    • (n+2) × 11.5 µL PPMV-1 Reaction Buffer

    • (n+2) × 11.5 µL PPMV-1 Premix

  3. Mix well and dispense 23 µL per PCR tube

  4. Add 2 µL of sample RNA, positiveあるいは negative control

  5. Final reaction volume: 25 µL


PCR Cycling Conditions:

StepTemperatureTime
Reverse Transcription50°C10 min
Pre-Denaturation95°C2 min
PCR Amplification (×40)95°C5 sec
59°C35 sec (FAM channel)

Compatible with ABI and other real-time PCR platforms. ROX dye optional.


Result Interpretation:

  • Positive: Ct ≤ 33 with clear “S” curve

  • Suspected: 33 < Ct < 38 → recommend retest

  • Negative: Ct ≥ 38 or no amplification curve

Control Requirements:

  • Positive Control: Ct < 28

  • Negative Control: Ct ≥ 38 or no Ct value


Precautions:

  1. Perform in separate work zones: RNA prep, reagent setup, and PCR amplification

  2. Wear gloves, a lab coat, and use separate pipettes and equipment for each area

  3. Avoid repeated freeze-thaw cycles; protect reagents from light

  4. For research use only; not intended for clinical diagnosis

  5. Disinfect the workspace and equipment regularly with alcohol, chlorine, or UV


🛒 Detect PPMV-1 early and effectively.
An essential tool for breeders, diagnostic labs, and avian disease researchers to safeguard flock health and reduce mortality risk from pigeon paramyxovirus outbreaks.

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