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PCR and ELISA vis-à-vis microscopy for detection of bovine anaplasmosis: a study on associated risk of an upcoming problem in North India.

Sharma A, Singla LD, Kaur P, Bal MS - ScientificWorldJournal (2015)

Bottom Line: Five false negative samples by msp1β PCR were reconfirmed for Anaplasma spp. targeting 16S rRNA gene.The sequence analysis showed the presence for A. marginale specific restriction site, indicating variation in the local strains of the organism resulting in no amplification with msp1β gene primers.All three districts of Submountain Zone are at higher risk indicating the impact of biotic and abiotic factors on the incidence of disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Parasitology, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab 141004, India.

ABSTRACT
This investigation demonstrates the status of bovine anaplasmosis caused by A. marginale in bovines from Submountain and Undulating Zone of Punjab. Out of 184 suspected animals, 25 (19.51%), 47 (31.71%), and 78 (68.75%) were positive by microscopy, indirect ELISA, and PCR assay, respectively. The microscopy showed 29% sensitivity and 99% specificity, while ELISA showed 32% sensitivity and 79% specificity in concordance with PCR assay. Five false negative samples by msp1β PCR were reconfirmed for Anaplasma spp. targeting 16S rRNA gene. The sequence analysis showed the presence for A. marginale specific restriction site, indicating variation in the local strains of the organism resulting in no amplification with msp1β gene primers. Of 82 samples positive by PCR, 57 were negative by ELISA indicating lower efficacy of ELISA to detect early anaplasmosis. The assessment of risk factor with results of PCR technique indicated that cattle (Odds ratio = 2.884), particularly those of age > 1 years (Odds ratio = 2.204) of district Pathankot (Odds ratio = 3.182) of Submountain Zone (Odds ratio = 2.086), were at high risk of anaplasmosis. All three districts of Submountain Zone are at higher risk indicating the impact of biotic and abiotic factors on the incidence of disease.

No MeSH data available.


Related in: MedlinePlus

Incidence of A. marginale among different animal groups under study as diagnosed by Blood Smear Examination (BSE), Enzyme Linked Immunosorbent Assay (ELISA), and Polymerase Chain Reaction (PCR) revealing cattle as the most exposed group.
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Related In: Results  -  Collection


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fig2: Incidence of A. marginale among different animal groups under study as diagnosed by Blood Smear Examination (BSE), Enzyme Linked Immunosorbent Assay (ELISA), and Polymerase Chain Reaction (PCR) revealing cattle as the most exposed group.

Mentions: Between the two species under study, cattle were shown to have higher susceptibility/exposure to the disease as revealed by serological (28.68%; 95% CI = 21.95–35.41%) as well as molecular test (49.61%; 95% CI = 11.23–35.82%). The incidence of disease varied significantly among the four animal groups with the PCR, highest being in cattle adults (58.11%) (95% CI = 48.41–67.80%) and lowest being in buffalo calves (23.57%) (95% CI = 11.23–35.82%) (Table 1, Figure 2). However, by microscopy and ELISA, there existed nonsignificant difference in the incidence of disease among the four animal groups. Cattle showed highest incidence of disease both by microscopy (18.92%) (95% CI = 11.22–26.61%) and ELISA (31.08%) (95% CI = 21.98–40.17%); however, study explicates the role of buffalo as the potent carrier animal by not exhibiting any clinical symptoms but still harbouring the infection. Cattle displayed a higher incidence of disease than buffaloes by all the three diagnostic techniques. Cattle were found to be more prone to anaplasmosis infection as compared to their calves. Similar trend was seen among buffaloes as revealed by PCR; however, the detection based on microscopy and ELISA depicted higher incidence of the disease in buffalo calves (Figure 2). Significant difference in the seroprevalence of A. marginale among different groups [9] may be influenced by factors such as race, age, and physiological and immunological status. It is noteworthy to mention that parasite inoculation rate by other biological vectors [10–12] and other sources of mechanical transmission, namely, blood-sucking dipterans and fomites [13] may also affect the enzootic stability of any geographic area when A. marginale is prevalent. Additionally, the lowest seropositivity (p < 0.05) seen in cattle aged up to 12 months can be explained by immune protection due to colostral antibodies and/or low rate of inoculation of A. marginale by the vectors as the animals in this age group have much lower contact with the vectors compared to older ones [14].


PCR and ELISA vis-à-vis microscopy for detection of bovine anaplasmosis: a study on associated risk of an upcoming problem in North India.

Sharma A, Singla LD, Kaur P, Bal MS - ScientificWorldJournal (2015)

Incidence of A. marginale among different animal groups under study as diagnosed by Blood Smear Examination (BSE), Enzyme Linked Immunosorbent Assay (ELISA), and Polymerase Chain Reaction (PCR) revealing cattle as the most exposed group.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4355815&req=5

fig2: Incidence of A. marginale among different animal groups under study as diagnosed by Blood Smear Examination (BSE), Enzyme Linked Immunosorbent Assay (ELISA), and Polymerase Chain Reaction (PCR) revealing cattle as the most exposed group.
Mentions: Between the two species under study, cattle were shown to have higher susceptibility/exposure to the disease as revealed by serological (28.68%; 95% CI = 21.95–35.41%) as well as molecular test (49.61%; 95% CI = 11.23–35.82%). The incidence of disease varied significantly among the four animal groups with the PCR, highest being in cattle adults (58.11%) (95% CI = 48.41–67.80%) and lowest being in buffalo calves (23.57%) (95% CI = 11.23–35.82%) (Table 1, Figure 2). However, by microscopy and ELISA, there existed nonsignificant difference in the incidence of disease among the four animal groups. Cattle showed highest incidence of disease both by microscopy (18.92%) (95% CI = 11.22–26.61%) and ELISA (31.08%) (95% CI = 21.98–40.17%); however, study explicates the role of buffalo as the potent carrier animal by not exhibiting any clinical symptoms but still harbouring the infection. Cattle displayed a higher incidence of disease than buffaloes by all the three diagnostic techniques. Cattle were found to be more prone to anaplasmosis infection as compared to their calves. Similar trend was seen among buffaloes as revealed by PCR; however, the detection based on microscopy and ELISA depicted higher incidence of the disease in buffalo calves (Figure 2). Significant difference in the seroprevalence of A. marginale among different groups [9] may be influenced by factors such as race, age, and physiological and immunological status. It is noteworthy to mention that parasite inoculation rate by other biological vectors [10–12] and other sources of mechanical transmission, namely, blood-sucking dipterans and fomites [13] may also affect the enzootic stability of any geographic area when A. marginale is prevalent. Additionally, the lowest seropositivity (p < 0.05) seen in cattle aged up to 12 months can be explained by immune protection due to colostral antibodies and/or low rate of inoculation of A. marginale by the vectors as the animals in this age group have much lower contact with the vectors compared to older ones [14].

Bottom Line: Five false negative samples by msp1β PCR were reconfirmed for Anaplasma spp. targeting 16S rRNA gene.The sequence analysis showed the presence for A. marginale specific restriction site, indicating variation in the local strains of the organism resulting in no amplification with msp1β gene primers.All three districts of Submountain Zone are at higher risk indicating the impact of biotic and abiotic factors on the incidence of disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Parasitology, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab 141004, India.

ABSTRACT
This investigation demonstrates the status of bovine anaplasmosis caused by A. marginale in bovines from Submountain and Undulating Zone of Punjab. Out of 184 suspected animals, 25 (19.51%), 47 (31.71%), and 78 (68.75%) were positive by microscopy, indirect ELISA, and PCR assay, respectively. The microscopy showed 29% sensitivity and 99% specificity, while ELISA showed 32% sensitivity and 79% specificity in concordance with PCR assay. Five false negative samples by msp1β PCR were reconfirmed for Anaplasma spp. targeting 16S rRNA gene. The sequence analysis showed the presence for A. marginale specific restriction site, indicating variation in the local strains of the organism resulting in no amplification with msp1β gene primers. Of 82 samples positive by PCR, 57 were negative by ELISA indicating lower efficacy of ELISA to detect early anaplasmosis. The assessment of risk factor with results of PCR technique indicated that cattle (Odds ratio = 2.884), particularly those of age > 1 years (Odds ratio = 2.204) of district Pathankot (Odds ratio = 3.182) of Submountain Zone (Odds ratio = 2.086), were at high risk of anaplasmosis. All three districts of Submountain Zone are at higher risk indicating the impact of biotic and abiotic factors on the incidence of disease.

No MeSH data available.


Related in: MedlinePlus