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A novel real-time PCR to detect Cetacean morbillivirus in Atlantic cetaceans
Kátia R Groch1, Sueli Akemi Taniwaki2, Cíntia Maria Favero1
1Laboratório de Patologia Comparada de Animais Selvagens, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brazil.
Abstract:
Cetacean morbillivirus (CeMV, family Paramyxoviridae) is a re-emergent pathogen associated with severe epizootic outbreaks causing high mortality among cetaceans worldwide. Recently, CeMV caused an unusual mortality event of Guiana dolphins (Sotalia guianensis) in Brazil. Partial sequence of the viral phosphoprotein (P) gene showed that the Guiana dolphin morbillivirus (GDMV) might represent a new lineage of CeMV. This study aimed to develop a molecular technique to detect the most common CeMV strains known to circulate in the Atlantic Ocean: GDMV, Dolphin morbillivirus (DMV) and Pilot-whale morbillivirus (PWMV). A sensible real-time reverse transcription polymerase chain reaction (RT-qPCR) method based on intercalating dye, targeting the P gene was described. This assay successfully detected GDMV, PWMV and DMV from field samples. Its performance was compared to a RT-qPCR method that specifically detects GDMV. Both assays had high sensibility and excellent intra- and inter-assay reproducibility. A total of 109 field samples from 32 Guiana dolphins were screened for CeMV by conventional RT-PCR in parallel with the RT-qPCR assay. The detection rate increased from 32% to 60% by use of the novel RT-qPCR. The RT-qPCR assay described herein allows rapid and sensitive detection of Atlantic CeMV strains, and is potentially suitable for screening of CeMV globally.
Insights
A new real-time PCR method rapidly detects Cetacean morbillivirus (CeMV) strains, including Guiana dolphin morbillivirus (GDMV), Dolphin morbillivirus (DMV), and Pilot-whale morbillivirus (PWMV). This sensitive assay improves detection rates in cetacean populations.
Area of Science:
- Veterinary Virology
- Marine Mammal Health
- Molecular Diagnostics
Background:
- Cetacean morbillivirus (CeMV) causes widespread, fatal epizootics in marine mammals.
- A recent Guiana dolphin mortality event in Brazil highlighted the need for better CeMV detection.
- Guiana dolphin morbillivirus (GDMV) may represent a distinct CeMV lineage.
Purpose of the Study:
- To develop a sensitive molecular diagnostic assay for common Atlantic CeMV strains.
- To target the phosphoprotein (P) gene for broad CeMV detection.
- To compare the novel assay's performance against existing methods.
Main Methods:
- Development of a real-time reverse transcription polymerase chain reaction (RT-qPCR) assay using intercalating dye.
- Targeting the conserved P gene of CeMV for detection.
- Validation using field samples and comparison with a GDMV-specific RT-qPCR and conventional RT-PCR.
Main Results:
- The novel RT-qPCR assay successfully detected GDMV, DMV, and PWMV in field samples.
- The assay demonstrated high sensitivity and excellent reproducibility.
- Utilizing the new RT-qPCR assay increased CeMV detection rates from 32% to 60% in Guiana dolphins.
Conclusions:
- A rapid, sensitive RT-qPCR assay for detecting Atlantic CeMV strains was established.
- This method enhances diagnostic capabilities for morbillivirus in cetaceans.
- The assay holds potential for global CeMV surveillance and screening efforts.
