Figure 3个  Table 3个
    • Serum
      number       		Sampling
      location       		Sampling
      date       		Patient
      sex       		Patient
      age (years)       		Clinical symptoms Traveling
      abroad history       		Mosquito
      bite
      #1 SGHS* 12/23/2016 F 50 Fever, chills, body ache,
      headache, joint pain, rash       		No Yes
      #2 SGHS 12/23/2016 F 45 Fever, chills, body ache, headache,
      joint pain, anorexia       		No Yes
      #3 SGHS 12/23/2016 F 50 Fever, chills, body ache, headache,
      joint pain, anorexia, rash       		No Yes
      #4 SGHS 12/23/2016 F 18 Fever, chills, body ache, headache,
      joint pain, anorexia, rash       		No Yes
      #5 SGHS 12/23/2016 F 40 Fever, chills, body ache, headache,
      joint pain       		No Yes
      #6 SGHS 12/23/2016 F 65 Fever, chills, body ache, headache,
      joint pain, rash       		No Yes
      #7 SGHS 01/05/2017 F 29 Fever, chills, body ache, headache,
      joint pain, rash       		No Yes
      #8 SGHS 01/05/2017 F 25 Fever, chills, body ache, headache,
      joint pain anorexia       		No Yes
      #9 SGHS 01/05/2017 M 15 Fever, chills, body ache, headache,
      joint pain, anorexia       		No Yes
      #10 SGHS 01/05/2017 M 14 Fever, chills, body ache, headache,
      joint pain, anorexia, rash       		No Yes
      Note: *SGHS: Sindh Government Hospital, Saudabad, Malir, Karachi.

      Table 1.  Information on 10 patients potentially infected with CHIKV in Pakistan

    • Serum specimen number Ct value
      (mean ± SD)       		Mean genomic RNA copy number (copies/μL)
      #1 19.69 ± 0.0283 1.1 × 107
      #2 33.52 ± 0.4243 1.2 × 103
      #3 22.40 ± 0.1626 1.8 × 106
      #4 21.40 ± 0.0919 3.6 × 106
      #5 28.84 ± 0.0141 2.6 × 104
      #6 19.14 ± 0.1768 1.6 × 107
      #7 21.08 ± 0.1414 4.4 × 106
      #8 Undetermined –
      #9 21.53 ± 0.0849 3.2 × 106
      #10 18.60 ± 0.0141 2.0 × 107
      Positive control 8.24 ± 0.03606 >109
      Negative control Undetermined –
      Note: The equation of the standard curve constructed for the SYBR Green real-time RT–PCR assay was: Y = –3.496X + 44.289 (R2 = 0.998; efficiency = 93.219%).

      Table 2.  Detection and quantification of CHIKV in 10 human serum specimens from Pakistan

    • Lineage Serum number/Location Strain GenBank ID Nucleotide/amino acid position in E1
      295/99 414/138 666/222 705/235 1030/344
      ECSA #01/Pakistan Pakistan-01 MF740875 G/E A/Q G/Q G/Q G/V
      #03/Pakistan Pakistan-03 MF740874 G/E A/Q G/Q G/Q A/I
      #04/Pakistan Pakistan-04 MF740876 G/E A/Q G/Q G/Q A/I
      #05/Pakistan Pakistan-05 MF740877 G/E A/Q G/Q C/H A/I
      #06/Pakistan Pakistan-06 MF740878 G/E G/Q G/Q G/Q A/I
      #07/Pakistan Pakistan-07 MF740879 A/K A/Q G/Q G/Q A/I
      #09/Pakistan Pakistan-09 MF740880 G/E A/Q G/Q G/Q A/I
      #10/Pakistan Pakistan-10 MF740881 G/E A/Q A/Q G/Q A/I
      –/India 119067 KY057363 G/E A/Q G/Q G/Q A/I
      Asian –/China CHIKV-JC2012 KC488650 G/E A/Q G/Q G/Q A/I
      West African –/Senegal 37997 AY726732 G/E A/Q G/Q G/Q G/V
      Note: The nucleotides and amino acids in bold are variable, including nonsynonymous substitutions, in CHIKV E1 from the ECSA lineage. Strains 119067, CHIKV-JC2012, and 37997 are representatives of the ECSA, Asian, and West African lineages, respectively.

      Table 3.  Variations in the E1 sequences of CHIKV strains from eight human serum samples