A method of simultaneously detecting Babesia species of B. microti, B. divergens, B. duncani, and/or B. venatorum in a sample, the method comprising: (a) performing an amplification step comprising contacting the sample with one or more set of primers for amplification of B. microti, B. divergens, B. duncani, and B. venatorum to produce an amplification product, if a target nucleic acid of B. microti, B. divergens, B. duncani, and/or B. venatorum is present in the sample; (b) performing a hybridization step comprising contacting the amplification product, if the target nucleic acid of B. microti, B. divergens, B. duncani, and/or B. venatorum is present in the sample, with one or more probes for hybridizing to the amplification product of B. microti, B. divergens, B. duncani, and B. venatorum; and (c) detecting the presence or absence of the amplification product, wherein the presence of the amplification product is indicative of the presence of B. microti, B. divergens, B. duncani, and/or B. venatorum in the sample, and wherein the absence of the amplification product is indicative of the absence of B. microti, B. divergens, B. duncani, and/or B. venatorum in the sample; and wherein the one or more set of primers and the one or more probes comprise a set of primers for amplification of B. microti comprising: (i) a first primer comprising the nucleic acid sequence of SEQ ID NO: 1, or a complement thereof; and (ii) a second primer comprising the nucleic acid sequence of SEQ ID NO:3, or a complement thereof; and a probe for hybridizing to the amplification product of B. microti comprising the nucleic acid sequence of SEQ ID NO:2, or a complement thereof; and a set of primers for amplification of B. divergens, B. duncani, and B. venatorum comprising: (i) a primer comprising the nucleic acid sequence of SEQ ID NO:4, or a complement thereof; (ii) one or more primers comprising a nucleic acid sequence of any one of a group consisting of SEQ ID NOs:6 and 7, or a complement thereof; and a probe for hybridizing to the amplification products of B. divergens, B. duncani, and B. venatorum comprising the nucleic acid sequence of SEQ ID NO:5, or a complement thereof. (a) performing an amplification step comprising contacting the sample with one or more set of primers for amplification of B. microti, B. divergens, B. duncani, and B. venatorum to produce an amplification product, if a target nucleic acid of B. microti, B. divergens, B. duncani, and/or B. venatorum is present in the sample; (b) performing a hybridization step comprising contacting the amplification product, if the target nucleic acid of B. microti, B. divergens, B. duncani, and/or B. venatorum is present in the sample, with one or more probes for hybridizing to the amplification product of B. microti, B. divergens, B. duncani, and B. venatorum; and (c) detecting the presence or absence of the amplification product, wherein the presence of the amplification product is indicative of the presence of B. microti, B. divergens, B. duncani, and/or B. venatorum in the sample, and wherein the absence of the amplification product is indicative of the absence of B. microti, B. divergens, B. duncani, and/or B. venatorum in the sample; and a set of primers for amplification of B. microti comprising: (i) a first primer comprising the nucleic acid sequence of SEQ ID NO: 1, or a complement thereof; and (ii) a second primer comprising the nucleic acid sequence of SEQ ID NO:3, or a complement thereof; and a probe for hybridizing to the amplification product of B. microti comprising the nucleic acid sequence of SEQ ID NO:2, or a complement thereof; and (i) a first primer comprising the nucleic acid sequence of SEQ ID NO: 1, or a complement thereof; and (ii) a second primer comprising the nucleic acid sequence of SEQ ID NO:3, or a complement thereof; and a set of primers for amplification of B. divergens, B. duncani, and B. venatorum comprising: (i) a primer comprising the nucleic acid sequence of SEQ ID NO:4, or a complement thereof; (ii) one or more primers comprising a nucleic acid sequence of any one of a group consisting of SEQ ID NOs:6 and 7, or a complement thereof; and a probe for hybridizing to the amplification products of B. divergens, B. duncani, and B. venatorum comprising the nucleic acid sequence of SEQ ID NO:5, or a complement thereof. (i) a primer comprising the nucleic acid sequence of SEQ ID NO:4, or a complement thereof; (ii) one or more primers comprising a nucleic acid sequence of any one of a group consisting of SEQ ID NOs:6 and 7, or a complement thereof; and