RESUMEN
Haemonchus contortus is one of the most significant strongylid nematodes infecting small ruminants, and it causes great economic losses to the livestock industry worldwide. Accurate diagnosis of H. contortus is crucial to control strategies. Traditional microscopic examinations are the most common methods for the diagnosis of H. contortus , but they are time-consuming and inaccurate. Molecular methods based on PCR are more accurate, but need expensive machines usually only used in the laboratory. Loop-mediated isothermal amplification (LAMP) is a rapid, simple, specific, and sensitive method that has been widely used to detect viruses, bacteria, and parasites. In the present study, a LAMP method targeting ribosomal ITS-2 gene for detection of the H. contortus in goat fecal samples has been established. The established LAMP method was H. contortus specific, and the sensitivity of LAMP was the same as that of the H. contortus species-specific PCR, with the lowest DNA level detected as being 1 pg. Examination of the clinical samples indicated that the positive rate of LAMP was higher than that of PCR, but no statistical difference was observed between LAMP and PCR (χ2 = 17.991, P = 0.053). In conclusion, a LAMP assay with a high specificity and a good sensitivity has been developed to detect H. contortus infection in goats. The established LAMP assay is useful for clinical diagnosis of H. contortus .
Asunto(s)
Heces/parasitología , Enfermedades de las Cabras/parasitología , Hemoncosis/veterinaria , Haemonchus/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico/métodos , Animales , Secuencia de Bases , Cartilla de ADN/química , ADN de Helmintos/química , ADN de Helmintos/aislamiento & purificación , Enfermedades de las Cabras/diagnóstico , Cabras , Hemoncosis/diagnóstico , Hemoncosis/parasitología , Haemonchus/genética , Técnicas de Amplificación de Ácido Nucleico/normas , Reacción en Cadena de la Polimerasa/veterinaria , Sensibilidad y Especificidad , Especificidad de la EspecieRESUMEN
UNLABELLED: Carbon dioxide (CO2 ) metabolically produced by humans has been widely used as a tracer gas for determining ventilation rates in occupied rooms. Among other necessities, the method requires good estimates of human CO2 generation rates. An empirically derived equation is widely used to calculate the CO2 generation rate. However, there are indications that this equation is not valid for young Chinese people. In this study, we measured the CO2 generation rate of 44 young Chinese people at two typical activity levels, quiet sitting and relaxed standing. We found that the commonly used empirical equation overpredicted CO2 generation rates, but could be corrected with a factor of 0.75 for Chinese females and of 0.85 for Chinese males. The variance for measured CO2 sitting was much smaller than for standing, and hence, we concluded that sitting yields more precise CO2 generation estimates. The relative contributions of sex, height, weight, and metabolic rate were analyzed. We concluded that the error in estimating metabolic rate is responsible for most of the difference in measured generation of CO2 from the empirical equation's predictions. PRACTICAL IMPLICATIONS: The tracer gas method using CO2 generated by people is widely used to calculate ventilation rate. However, the empirically derived equation that is normally used to estimate CO2 generation rate is not suitable for young Chinese people at rest. To estimate the CO2 generation rate in Chinese people under low-activity conditions, the empirical equation should be multiplied by correction factors of 0.75 and 0.85 for females and males, respectively.