Detection of hepatopancreatic parvovirus (HPV) infection in Penaeus monodon using PCR-ELISA.
A rapid and sensitive PCR-ELISA has been developed for detection of hepatopancreatic parvovirus (HPV) in Penaeus monodon. The specific primer set amplified 156 bp fragment and could detect as a little as 0.01 fg of purified HPV DNA which equivalent to three viral particles. No cross-reactivity was observed when nucleic acid templates from white spot syndrome virus, yellow-head virus, monodon baculovirus and shrimp were tested. The crude DNA simple prepared from hepatopancreas can be used as DNA template and provide a favorable result. Using this technique for detection of HPV infection in 87 carrier shrimps revealed the higher sensitivity and efficiency of detection when compared to histological examination and conventional PCR. Sixty-two percent infection was detected by PCR-ELISA from samples with HPV negative diagnosed by histological examination. Therefore, this sensitive and specific method is promisingly useful for early detection of HPV infection in broodstock, carriers and for ex situ application where large numbers of samples can be analyzed simultaneously.
Sukhumsirichart
W
W
Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand. wasanas@psm.swu.ac.th
Kiatpathomchai
W
W
Wongteerasupaya
C
C
Withyachumnarnkul
B
B
Flegel
T W
TW
Boonseang
V
V
Panyim
S
S
D000818:Animals; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D004064:Digestive System / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D010321:Parvoviridae / Q000235:genetics / Q000302:isolation & purification*; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary / Q000821:virology*; D033561:Penaeidae / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral