03253nas a2200301   4500008004100000022001400041245015900055210006900214260000800283300001100291490000800302520224400310653002002554653000902574653001402583653000802597653001702605653001402622100002702636700002302663700002102686700001902707700002502726700002402751700002802775700002202803856012602825       2019                        eng d  a0014-489400aMitochondrial cytochrome oxidase C subunit III (cox3) gene as a sensitive and specific target for molecular detection of Babesia gibsoni infection in dogs0 aMitochondrial cytochrome oxidase C subunit III cox3 gene as a se  cNOV  a1077710 v2063 a<p>A PCR targeting mitochondrial cytochrome oxidase subunit III (cox3) for molecular detection of Babesia gibsoni infection in dogs has been developed in this study. Fifty blood samples from suspected clinical cases from dogs, brought to the veterinary college clinics, were examined for presence of B. gibsoni using conventional diagnosis by microscopic examination of Giemsa stained thin blood smears. In addition, species specific PCRs targeting ITS-1 region (BgITS-1 PCR) and nested PCR targeting 18S ribosomal RNA gene (Bg18SnPCR) were carried out. A 634 bp PCR fragment of B. gibsoni cox3 gene was amplified in positive samples from three geographical locations of Satara, Wai and Pune in Maharashtra state of India. From analysis of the sequence of the B. gibsoni cox3 gene, we found that the Indian isolate had 96-98% similarity to the isolate from Japan and China. Post sequencing, de-novo diagnostic primer pair for species specific amplification of 164 bp fragment of B. gibsoni cox3 was designed and the PCR was standardized. The diagnostic results of de-novo Bgcox3 PCR were compared with BgITS-1 PCR and Bg18S nPCR. Thin blood smears detected 22% (11/50) samples positive for small form of Babesia species. The BgITS-1 PCR detected 25% samples (15/50) as positive and Bg18S nPCR detected 80% (40/50) B. gibsoni positive samples. The de-novo Bgcox3 PCR detected 66% (33/50) samples positive for B. gibsoni (at 95% CI). The analytical sensitivity of cox3 PCR was evaluated as 0.000003% parasitaemia or 09 parasites in 100 mu l of blood. The de-novo diagnostic cox3 PCR did not cross react with control positive DNA from other haemoprotozoa and rickettsia like B. vogeli, Hepatozoon canis, Trypanosoma evansi, Ehrlichia cants and Anaplasma platys. Statistically, cox3 PCR had better diagnostic efficiency than ITS-1 PCR in terms of sensitivity (p = 0.0006). No statistically significant difference between results of cox3 PCR and 18S nPCR was observed (p = 0.1760). Kappa values estimated for each test pair showed fair to moderate agreement between the observations. Specificity of Bgcox3 PCR was 100% when compared with microscopy or BgITS-1 PCR. Sensitivity of Bgcox3 PCR was 100% when compared with that of Bg18S nPCR.</p>
10aBabesia gibsoni10acox310aDiagnosis10aDog10amitochondria10amolecular1 aBetgiri, Aishwarya, A.1 aJadhav, Sameer, N.1 aPawde, Mrunalini1 aShukla, Anurag1 aMote, Chandrashekhar1 aPawar, Prashant, D.1 aShanmugam, Dhanasekaran1 aKundu, Krishnendu  uhttps://library.ncl.res.in/content/mitochondrial-cytochrome-oxidase-c-subunit-iii-cox3-gene-sensitive-and-specific-target02085nas a2200253   4500008004100000022001400041245009600055210006900151260000800220300001400228490000800242520125000250100001801500700001901518700002001537700002001557700002101577700002101598700002101619700002801640700002601668700002201694856011501716       2022                        eng d  a0002-963700aCase report: a fatal case of babesiosis in a splenectomized male patient from Western India0 aCase report a fatal case of babesiosis in a splenectomized male   cMAY  a1421-14250 v1063 a<p>
	Human babesiosis is a rare disease, caused by Babesia species and commonly transmitted by tick bite. Although human babesiosis is known to be asymptomatic in immunocompetent hosts, clinical cases of severe babesiosis have been reported from splenectomized or immunocompromised individuals. To our knowledge, only one case of human babesiosis in India has been previously reported. Here, we report a case of severe babesiosis with high parasitemia (similar to 70%) in a 30-year-old asplenic farmer. The patient presented with fever, yellowish discoloration of skin, oliguria, and anemia; he eventually developed multiorgan failure syndrome and died. Peripheral blood films were prepared and used to confirm the presence of piroplasms by microscopy. Total DNA isolated from blood was used for 18S ribosomal RNA gene fragment amplification by polymerase chain reaction, which was subjected to Sanger sequencing. Although 18S sequence indicated that the Babesia species infecting the patient was similar to that of other Babesia species originating from wild mammals, species identification could not be done. Phylogenetic analysis revealed that the patientderived pathogen is distinct because it forms a separate clade in the cladogram.</p>
1 aGodbole, Ravi1 aGaur, Avantika1 aNayar, Priyanka1 aKiruthiga, Kala1 aD'Costa, Pradeep1 aManchanda, Rumma1 aKhilari, Ajinkya1 aShanmugam, Dhanasekaran1 aMuglikar, Kalpana, D.1 aKundu, Krishnendu  uhttps://library.ncl.res.in/content/case-report-fatal-case-babesiosis-splenectomized-male-patient-western-india