Multiplexed-tandem PCR (MT-PCR) assay to detect and differentiate gastrointestinal nematodes of alpacas

• Published in: Parasites & vectors Vol. 11; no. 1; p. 370
• Main Authors: Rashid, Mohammed H, Gebrekidan, Hagos, Jabbar, Abdul
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 28.06.2018; BioMed Central; BMC
• Subjects: Alpaca; Alpacas; Analysis; Animals; Assaying; Australia; Australia - epidemiology; Automation; Camelidae; Camelids, New World - parasitology; Cooperia; Data Accuracy; Deoxyribonucleic acid; Diagnosis; DNA; Economic impact; Eggs; Epidemiology; Feces; Feces - parasitology; Gastrointestinal diseases; Gastrointestinal Tract - parasitology; Health aspects; Identification; Infections; Intestinal parasites; Larvae; Leather; Llamas; Meat; Methods; Morphology; Multiplex Polymerase Chain Reaction - methods; Multiplexing; Nematoda - genetics; Nematoda - isolation & purification; Nematode Infections - diagnosis; Nematode Infections - epidemiology; Nematode Infections - parasitology; Nematode Infections - veterinary; Nematodes; Nucleotide sequence; Ostertagia - genetics; Ostertagia - isolation & purification; Ostertagia ostertagi; Parasites; PCR; Polymerase chain reaction; Sensitivity and Specificity; Sheep; Sheep Diseases - diagnosis; Sheep Diseases - epidemiology; Sheep Diseases - parasitology; Short Report; South American camelids; Teladorsagia circumcincta; Australia; Nematodes; Alpacas; Llamas; South American camelids; Australia; PCR
• ISSN: 1756-3305; 1756-3305
• DOI: 10.1186/s13071-018-2963-9

Gastrointestinal nematodes (GINs) frequently infect South American camelids (alpacas and llamas) and cause economic losses due to reduced production of fiber, meat and/or leather. Our knowledge about the epidemiology and diagnosis of GINs in llamas and alpacas is limited, and reliable keys for the identification of the third-stage larvae (L3s) of some common nematodes (such as Camelostrogylus mentulatus) that infect alpacas and llamas remain undescribed. In this study, we modified two existing semi-quantitative multiplexed-tandem (MT)-PCR assays, originally developed for the GINs of sheep and cattle, to reliably detect and differentiate the common genera/species of GINs in the faeces of alpacas. Following the establishment of the MT-PCR assay using positive and negative control samples, alpaca faecal samples were tested to validate the assay to detect and differentiate nematode genera/species, including C. mentulatus, Cooperia spp., Haemonchus spp., Oesophagostomum spp., Ostertagia ostertagi, Teladorsagia circumcincta and Trichostrongylus spp. Sequencing of the MT-PCR products demonstrated specific (100%) amplification of the target nematode genera/species. Additionally, a comparison of results of the MT-PCR assay and the morphological identification of adult worms collected from the same 35 alpacas revealed that there was a good agreement (37-94%) between the two methods. However, some discrepancies were observed between the results of the MT-PCR assay and the morphological identification of adult worms. The MT-PCR platform is an accurate, sensitive and rapid method for the diagnosis of GINs in alpacas, and it can be used as a substitute to larval culture to identify common nematodes in the faeces of alpacas and llamas.