diff --git a/tests/test_bcftools_validation.py b/tests/test_bcftools_validation.py
index d5dd564..2d63962 100644
--- a/tests/test_bcftools_validation.py
+++ b/tests/test_bcftools_validation.py
@@ -61,7 +61,11 @@ def run_vcztools(args: str) -> str:
         ("view --no-version -s NA00001", "sample.vcf.gz"),
         ("view --no-version -s NA00001,NA00003", "sample.vcf.gz"),
         ("view --no-version -s HG00096", "1kg_2020_chrM.vcf.gz"),
-        ("view --no-version -s '' --force-samples", "sample.vcf.gz")
+        ("view --no-version -s '' --force-samples", "sample.vcf.gz"),
+        ("view --no-version -s ^NA00001", "sample.vcf.gz"),
+        ("view --no-version -s ^NA00003,NA00002", "sample.vcf.gz"),
+        ("view --no-version -s ^NA00003,NA00002,NA00003", "sample.vcf.gz"),
+        ("view --no-version -S ^tests/data/txt/samples.txt", "sample.vcf.gz"),
     ]
 )
 # fmt: on
diff --git a/tests/test_cli.py b/tests/test_cli.py
index d0f1c38..a955e5d 100644
--- a/tests/test_cli.py
+++ b/tests/test_cli.py
@@ -7,21 +7,32 @@
 from tests.utils import vcz_path_cache
 
 
-def test_version_header():
+@pytest.fixture()
+def vcz_path():
     vcf_path = pathlib.Path("tests/data/vcf/sample.vcf.gz")
-    vcz_path = vcz_path_cache(vcf_path)
+    return vcz_path_cache(vcf_path)
 
+
+def test_version_header(vcz_path):
     output = run_vcztools(f"view {vcz_path}")
     assert output.find("##vcztools_viewCommand=") >= 0
     assert output.find("Date=") >= 0
 
 
-def test_view_bad_output(tmp_path):
-    vcf_path = pathlib.Path("tests/data/vcf/sample.vcf.gz")
-    vcz_path = vcz_path_cache(vcf_path)
+def test_view_bad_output(tmp_path, vcz_path):
     bad_output = tmp_path / "output.vcf.gz"
 
     with pytest.raises(
         ValueError, match=re.escape("Output file extension must be .vcf, got: .gz")
     ):
         run_vcztools(f"view --no-version {vcz_path} -o {bad_output}")
+
+
+def test_excluding_and_including_samples(vcz_path):
+    samples_file_path = pathlib.Path("tests/data/txt/samples.txt")
+    error_message = re.escape("vcztools does not support combining -s and -S")
+
+    with pytest.raises(AssertionError, match=error_message):
+        run_vcztools(f"view {vcz_path} -s NA00001 -S ^{samples_file_path}")
+    with pytest.raises(AssertionError, match=error_message):
+        run_vcztools(f"view {vcz_path} -s ^NA00001 -S {samples_file_path}")
diff --git a/tests/test_vcf_writer.py b/tests/test_vcf_writer.py
index 0df9c1c..57f7845 100644
--- a/tests/test_vcf_writer.py
+++ b/tests/test_vcf_writer.py
@@ -145,14 +145,17 @@ def test_write_vcf__regions(tmp_path, regions, targets,
         assert variant.POS == pos
 
 @pytest.mark.parametrize(
-    ("samples", "expected_genotypes"),
+    ("samples", "expected_samples", "expected_genotypes"),
     [
-        ("NA00001", [[0, 0, True]]),
-        ("NA00001,NA00003", [[0, 0, True], [0, 1, False]]),
-        ("NA00003,NA00001", [[0, 1, False], [0, 0, True]]),
+        ("NA00001", ["NA00001"], [[0, 0, True]]),
+        ("NA00001,NA00003", ["NA00001", "NA00003"], [[0, 0, True], [0, 1, False]]),
+        ("NA00003,NA00001", ["NA00003", "NA00001"], [[0, 1, False], [0, 0, True]]),
+        ("^NA00002", ["NA00001", "NA00003"], [[0, 0, True], [0, 1, False]]),
+        ("^NA00003,NA00002", ["NA00001"], [[0, 0, True]]),
+        ("^NA00003,NA00002,NA00003", ["NA00001"], [[0, 0, True]]),
     ]
 )
-def test_write_vcf__samples(tmp_path, samples, expected_genotypes):
+def test_write_vcf__samples(tmp_path, samples, expected_samples, expected_genotypes):
     original = pathlib.Path("tests/data/vcf") / "sample.vcf.gz"
     vcz = vcz_path_cache(original)
     output = tmp_path.joinpath("output.vcf")
@@ -161,7 +164,7 @@ def test_write_vcf__samples(tmp_path, samples, expected_genotypes):
 
     v = VCF(output)
 
-    assert v.samples == samples.split(",")
+    assert v.samples == expected_samples
 
     variant = next(v)
 
diff --git a/vcztools/cli.py b/vcztools/cli.py
index 049de22..99e5a30 100644
--- a/vcztools/cli.py
+++ b/vcztools/cli.py
@@ -149,8 +149,15 @@ def view(
         raise ValueError(f"Output file extension must be .vcf, got: .{split[-1]}")
 
     if samples_file:
+        assert not samples, "vcztools does not support combining -s and -S"
+
+        samples = ""
+        exclude_samples_file = samples_file.startswith("^")
+        samples_file = samples_file.lstrip("^")
+
         with open(samples_file) as file:
-            samples = samples or ""
+            if exclude_samples_file:
+                samples = "^" + samples
             samples += ",".join(line.strip() for line in file.readlines())
 
     # TODO: use no_update when fixing https://github.com/sgkit-dev/vcztools/issues/75
diff --git a/vcztools/vcf_writer.py b/vcztools/vcf_writer.py
index d383210..ce6d50d 100644
--- a/vcztools/vcf_writer.py
+++ b/vcztools/vcf_writer.py
@@ -153,10 +153,18 @@ def write_vcf(
             samples_selection = None
         else:
             all_samples = root["sample_id"][:]
+            exclude_samples = samples.startswith("^")
+            samples = samples.lstrip("^")
             sample_ids = np.array(samples.split(","))
             if np.all(sample_ids == np.array("")):
                 sample_ids = np.empty((0,))
+
             samples_selection = search(all_samples, sample_ids)
+            if exclude_samples:
+                samples_selection = np.setdiff1d(
+                    np.arange(all_samples.size), samples_selection
+                )
+                sample_ids = all_samples[samples_selection]
 
         if not no_header and vcf_header is None:
             if "vcf_header" in root.attrs: